Antibody molecules to TIM-3 and uses thereof

ABSTRACT

Antibody molecules that specifically bind to TIM-3 are disclosed. The anti-TIM-3 antibody molecules can be used to treat, prevent and/or diagnose immune, cancerous, or infectious conditions and/or disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. application Ser.No. 14/610,837, filed Jan. 30, 2015, now U.S. Pat. No. 9,605,070, whichclaims the benefit of U.S. Provisional Application No. 61/934,469, filedJan. 31, 2014, and U.S. Provisional Application No. 62/094,912, filedDec. 19, 2014. The contents of the aforementioned applications arehereby incorporated by reference in their entirety.

GOVERNMENT SUPPORT

This invention was made with government support under grant numbers P01AI054456 and R01 AI089955 awarded by The National Institutes of Health.The government has certain rights in the invention.

BACKGROUND

Activation of naive CD4+ T helper cells results in the development of atleast two distinct effector populations, Th1 cells and Th2 cells. SeeU.S. Pat. No. 7,470,428, Mosmann T R et al. (1986) J Immunol136:2348-57; Mosmann T R et al. (1996) Immunol Today 17:138-46; Abbas AK et al. (1996) Nature 383:787-793. Th1 cells produce cytokines (e.g.,interferon gamma, interleukin-2, tumor necrosis factor alpha, andlymphotoxin) which are commonly associated with cell-mediated immuneresponses against intracellular pathogens, delayed-type hypersensitivityreactions (Sher A et al. (1992) Annu Rev Immunol 10:385-409), andinduction of organ-specific autoimmune diseases. Liblau R S et al.(1995) Immunol Today 16:34-38. Th2 cells produce cytokines (e.g., IL-4,IL-10, and IL-13) that are crucial for control of extracellularhelminthic infections and promote atopic and allergic diseases. Sher Aet al. (1992) Annu Rev Immunol 10:385-409. In addition to their distinctroles in disease, the Th1 and Th2 cells cross-regulate each other'sexpansion and functions. Thus, preferential induction of Th2 cellsinhibits autoimmune diseases (Kuchroo V K et al. (1995) Cell 80:707-18;Nicholson L B et al. (1995) Immunity 3:397-405), and predominantinduction of Th1 cells can regulate induction of asthma, atopy andallergies. Lack G et al. (1994) J Immunol 152:2546-54; Hofstra C L etal. (1998) J Immunol 161:5054-60.

TIM-3 is a transmembrane receptor protein that is expressed, e.g., onTh1 (T helper 1) CD4+ cells and cytotoxic CD8+ T cells that secreteIFN-γ. TIM-3 is generally not expressed on naïve T cells but ratherupregulated on activated, effector T cells. TIM-3 has a role inregulating immunity and tolerance in vivo (see Hastings et al., Eur JImmunol. 2009 September; 39(9):2492-501). There is a need in the art fornew molecules that regulate TIM-3 function and the function of TIM-3expressing cells.

SUMMARY

Disclosed herein are antibody molecules that bind to TIM-3 (T-cellimmunoglobulin domain and mucin domain 3) with high affinity andspecificity. Nucleic acid molecules encoding the antibody molecules,expression vectors, host cells and methods for making the antibodymolecules are also provided. Immunoconjugates, multi- or bispecificantibody molecules and pharmaceutical compositions comprising theantibody molecules are also provided. The anti-TIM-3 antibody moleculesdisclosed herein can be used (alone or in combination with other agentsor therapeutic modalities) to treat, prevent and/or diagnose immunedisorders, cancer, infectious disease, Crohn's disease, sepsis, SIRS(Systemic Inflammatory Response Syndrome), and glomerulonephritis. Thus,compositions and methods for detecting TIM-3, as well as methods fortreating various disorders, including cancer and immune disorders usingthe anti-TIM-3 antibody molecules are disclosed herein.

Accordingly, in certain aspects, this disclosure provides an antibodymolecule (e.g., an isolated or recombinant antibody molecule) having oneor more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 orall) of the following properties (a), (b), (c), (d), (e), (f), (g), (h),(i), (j), (k), (l), (m), (n), (o), (p) or (q):

-   -   (a) binds to TIM-3, e.g., human TIM-3, with high affinity, e.g.,        with a dissociation constant (K_(D)) of less than about 100 nM,        typically about 10 nM, and more typically, about 1-0.1 nM or        stronger, e.g., less than about 0.2, 0.16, 0.15, 0.1, 0.075,        0.05, or 0.042 nM,    -   (b) binds substantially to a non-human primate TIM-3, e.g.,        cynomolgus TIM-3, with a dissociation constant (K_(D)) of less        than about 100 nM, typically about 10 nM, and more typically,        about 3-0.3 nM or stronger, e.g., 1-0.1 nM or stronger, e.g.,        less than about 1 nM, 0.75 nM, or 0.68 nM,    -   (c) inhibits binding of TIM-3 to a TIM-3 ligand, e.g.,        phosphatidylserine (PtdSer), HMGB1, or CEACAM-1,    -   (d) enhances IFN-gamma and/or TNF-alpha secretion and/or        proliferation in T cells, e.g., CD4+ or CD8+ T cells, e.g., in        CD4+ T cells that were stimulated with anti-CD3/CD28 in the        presence of IL-12 or in T cell-DC autologous culture assays with        anti-CD3/CD28 stimulation,    -   (e) enhances cytotoxic NK (natural killer) cell activity against        a target cell (e.g., K562 cells), e.g., in an in vitro assay,    -   (f) enhances capacity of macrophages or antigen presenting cells        to stimulate a T cell response, e.g., increasing IL-12 secretion        of antigen presenting cells,    -   (g) binds specifically to an epitope on TIM-3, e.g., the same or        similar epitope as the epitope recognized by an antibody        molecule described herein, e.g., a murine or humanized        anti-TIM-3 antibody molecule as described herein, e.g., an        antibody molecule of Tables 1-4,    -   (h) shows the same or similar binding affinity or specificity,        or both, as an antibody molecule of Tables 1-4,    -   (i) shows the same or similar binding affinity or specificity,        or both, as an antibody molecule (e.g., an heavy chain variable        region and light chain variable region) described in Tables 1-4,    -   (j) shows the same or similar binding affinity or specificity,        or both, as an antibody molecule (e.g., an heavy chain variable        region and light chain variable region) comprising an amino acid        sequence shown in Tables 1-4,    -   (k) inhibits, e.g., competitively inhibits, the binding of a        second antibody molecule to TIM-3 wherein the second antibody        molecule is an antibody molecule described herein, e.g., an        antibody molecule chosen from Tables 1-4,    -   (l) binds the same (or substantially the same) or an overlapping        (or substantially overlapping) epitope with a second antibody        molecule to TIM-3, wherein the second antibody molecule is an        antibody molecule described herein, e.g., an antibody molecule        chosen from Tables 1-4,    -   (m) competes for binding, and/or binds the same (or        substantially the same) or overlapping (or substantially        overlapping) epitope, with a second antibody molecule to TIM-3,        wherein the second antibody molecule is an antibody molecule        described herein, e.g., an antibody molecule chosen from Tables        1-4, e.g., as determined by the methods described in Example 11,    -   (n) has one or more biological properties of an antibody        molecule described herein, e.g., an antibody molecule chosen        from Tables 1-4,    -   (o) has one or more pharmacokinetic properties of an antibody        molecule described herein, e.g., an antibody molecule chosen        from Tables 1-4,    -   (p) modulates (e.g., enhances or inhibits) one or more        activities of TIM-3, e.g., results in one or more of: enhancing        IFN-gamma and/or TNF-alpha secretion in T cells; enhancing        proliferation in T cells, e.g., CD4+ or CD8+ T cells; enhancing        NK cell cytotoxic activity; reducing suppressor activity of        regulatory T cells (Tregs); or increasing immune stimulation        properties of macrophages and/or antigen presenting cells, e.g.,        increasing cytokine secretion, e.g., IL-12 secretion; or    -   (q) binds to one or more residues within: the two residues        adjacent to the N-terminus of the A strand (residues Val24 and        Glu25 in human TIM-3), the BC loop, the CC′ loop, the F strand,        the FG loop, and the G strand of TIM-3, or one or more residues        within a combination of two, three, four, five or all of: the        two residues adjacent to the N-terminus of the A strand        (residues Val24 and Glu25 in human TIM-3), the BC loop, the CC′        loop, the F strand, the FG loop, and the G strand of TIM-3,        e.g., wherein the binding is assayed using ELISA or Biacore.

In some embodiments, the antibody molecule binds to TIM-3 with highaffinity, e.g., with a K_(D) that is at least about 10%, 20%, 30%, 40%,50%, 60%, 70%, 80% or 90% lower than the K_(D) of a murine anti-TIM-3antibody molecule, e.g., a murine anti-TIM-3 antibody molecule describedherein.

In some embodiments, the expression level of the anti-TIM-3 antibodymolecule is higher, e.g., at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9or 10-fold higher, than the expression level of a murine antibodymolecule, e.g., a murine or chimeric anti-TIM-3 antibody moleculedescribed herein. In some embodiments, the antibody molecule isexpressed in mammalian cells, e.g., rodent cells.

In some embodiments, the anti-TIM-3 antibody molecule reduces one ormore activities of TIM-3 with an IC50 (concentration at 50% inhibition)that is lower, e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%,80% or 90% lower, than the IC50 of a murine anti-TIM-3 antibodymolecule, e.g., a murine anti-TIM-3 antibody molecule described herein.In some embodiments, the TIM-3 activity is the binding of TIM-3 to oneor more (e.g., one, two, three, four or all) of the TIM-3 ligandsdescribed herein, e.g., one, two or more (all) of PtdSer, CEACAM-1, orHMGB1.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, or more continuous or discontinuous (e.g., noncontiguous)amino acid residues chosen from Val24, Glu25, Thr41, Gly56, Ala57,Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125,Lys126, and/or Leu127.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous ordiscontinuous (e.g., noncontiguous) amino acid residues chosen fromVal24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59,Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122,Phe123, Asn124, Leu125, Lys126, Leu127, and/or Val128, e.g., as detailedin Table 13.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous ordiscontinuous (e.g., noncontiguous) amino acid residues chosen fromGlu23, Val24, Glu25, Tyr26, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46,Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56,Ala57, Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124,Leu125, Lys126 and/or Leu127.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous ordiscontinuous (e.g., noncontiguous) amino acid residues chosen fromVal24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Pro42, Ala43, Ala44, Pro45,Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55,Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107,Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127,and/or Val128.

In other embodiments, the anti-TIM-3 antibody molecule competes withCEACAM-1 for binding to TIM-3. In one embodiment, the anti-TIM-3antibody molecule interacts, e.g., binds to, one, two, or more (all) ofCys58, Asn119 and Lys122 of TIM-3, e.g., displaces or competes CEACAM-1for binding to these residues. In one embodiment, the anti-TIM-3antibody molecule reduces or blocks the formation of a hydrogen bondbetween Lys122 of TIM-3 and Asn42 of CEACAM-1, e.g., by at least about10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, compared to the formationof a hydrogen bond between between Lys122 of TIM-3 and Asn42 of CEACAM-1in the absence of the anti-TIM-3 antibody molecule.

In another embodiment, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a PtdSer-binding loop of TIM-3. In one embodiment, theanti-TIM-3 antibody molecule interacts with, e.g., binds to, at leasttwo PtdSer-binding loops of TIM-3, e.g., the FG loop and CC′ loop ofTIM-3 (e.g., a metal ion-dependent ligand binding site (MILIBS)). Forexample, the carboxyl group of PtdSer can bind to the CC′ loop of TIM-3and the amino group of PtdSer can bind to the FG loop of TIM-3. In oneembodiment, the anti-TIM-3 antibody molecule reduces or preventsPtdSer-mediated membrane penetration of TIM-3.

In another embodiment, the anti-TIM-3 antibody molecule competes withHMGB1 for binding to TIM-3. E.g., it reduces binding of HMGB1 to residue62 of TIM-3 (Q in mouse, E in human TIM-3), e.g., by at least about 10%,20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, compared to the binding ofHMGB1 to residue 62 of TIM-3 in the absence of the anti-TIM-3 antibodymolecule.

In yet another embodiment, the anti-TIM-3 antibody molecule does notcompete with a Galectin-9 (Gal-9) ligand for binding to TIM-3.

In some embodiments, the anti-TIM-3 antibody molecule has improvedstability, e.g., at least about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 or10-fold more stable in vivo or in vitro, than a murine or humanizedanti-TIM-3 antibody molecule, e.g., a murine or humanized anti-TIM-3antibody molecule described herein.

In some embodiments, the anti-TIM-3 antibody molecule comprises at leastone antigen-binding region, e.g., a variable region or anantigen-binding fragment thereof, from an antibody described herein,e.g., an antibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4; or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences.

In certain embodiments, the anti-TIM-3 antibody molecule comprises atleast one, two, three, or four variable regions from an antibodydescribed herein, e.g., an antibody chosen from any of ABTIM3,ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05,ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10,ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15,ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20,ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4;or encoded by the nucleotide sequence in Tables 1-4; or a sequencesubstantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%,98%, 99% or higher identical) to any of the aforesaid sequences.

In some embodiments, the anti-TIM-3 antibody molecule comprises at leastone or two heavy chain variable regions from an antibody describedherein, e.g., an antibody chosen from any of ABTIM3, ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4; or encoded bythe nucleotide sequence in Tables 1-4; or a sequence substantiallyidentical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% orhigher identical) to any of the aforesaid sequences.

In certain embodiments, the anti-TIM-3 antibody molecule comprises atleast one or two light chain variable regions from an antibody describedherein, e.g., an antibody chosen from any of ABTIM3, ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4; or encoded bythe nucleotide sequence in Tables 1-4; or a sequence substantiallyidentical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% orhigher identical) to any of the aforesaid sequences.

In one embodiment, the anti-TIM-3 antibody molecule includes a heavychain constant region for an IgG4, e.g., a human IgG4. In anotherembodiment, the human IgG4 includes a substitution (e.g., a Ser to Prosubstitution) at position 228 according to EU numbering or at position108 of SEQ ID NO: 108 or 110. In still another embodiment, theanti-TIM-3 antibody molecule includes a heavy chain constant region foran IgG1, e.g., a human IgG1. In one embodiment, the human IgG1 includesa substitution (e.g., an Asn to Ala substitution) at position 297according to EU numbering or at position 180 of SEQ ID NO: 112. In oneembodiment, the human IgG1 includes a substitution (e.g., an Asp to Alasubstitution) at position 265 according to EU numbering or at position148 of SEQ ID NO: 113, a substitution (e.g., a Pro to Ala substitution)at position 329 according to EU numbering or at position 212 of SEQ IDNO: 113, or both. In one embodiment, the human IgG1 includes asubstitution (e.g., a Leu to Ala substitution) at position 234 accordingto EU numbering or at position 117 of SEQ ID NO: 114, a substitution(e.g., a Leu to Ala substitution) at position 235 according to EUnumbering or at position 118 of SEQ ID NO: 114, or both. In oneembodiment, the heavy chain constant region comprises an amino sequenceset forth in Table 1-5, or a sequence substantially identical (e.g., atleast 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical)thereto.

In yet another embodiment, the anti-TIM-3 antibody molecule includes akappa light chain constant region, e.g., a human kappa light chainconstant region. In one embodiment, the light chain constant regioncomprises an amino sequence set forth in Table 1-5, or a sequencesubstantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%,98%, 99% or higher identical) thereto.

In another embodiment, the anti-TIM-3 antibody molecule includes a heavychain constant region for an IgG4, e.g., a human IgG4, and a kappa lightchain constant region, e.g., a human kappa light chain constant region,e.g., a heavy and light chain constant region comprising an aminosequence set forth in Table 1-5, or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) thereto. In yet another embodiment, the anti-TIM-3 antibodymolecule includes a heavy chain constant region for an IgG1, e.g., ahuman IgG1, and a kappa light chain constant region, e.g., a human kappalight chain constant region, e.g., a heavy and light chain constantregion comprising an amino sequence set forth in Table 1-5, or asequence substantially identical (e.g., at least 80%, 85%, 90%, 92%,95%, 97%, 98%, 99% or higher identical) thereto. In one embodiment, thehuman IgG1 includes a substitution at position 297 according to EUnumbering (e.g., an Asn to Ala substitution). In one embodiment, thehuman IgG1 includes a substitution at position 265 according to EUnumbering, a substitution at position 329 according to EU numbering, orboth (e.g., an Asp to Ala substitution at position 265 and/or a Pro toAla substitution at position 329). In one embodiment, the human IgG1includes a substitution at position 234 according to EU numbering, asubstitution at position 235 according to EU numbering, or both (e.g., aLeu to Ala substitution at position 234 and/or a Leu to Ala substitutionat position 235).

In another embodiment, the anti-TIM-3 antibody molecule includes a heavychain variable domain and a constant region, a light chain variabledomain and a constant region, or both, comprising the amino acidsequence of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; oras described in Tables 1-4; or encoded by the nucleotide sequence inTables 1-4; or a sequence substantially identical (e.g., at least 80%,85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of theaforesaid sequences.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three complementarity determining regions (CDRs) from aheavy chain variable region of an antibody described herein, e.g., anantibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4, or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences.

In some embodiments, the anti-TIM-3 antibody molecule comprises at leastone, two, or three complementarity determining regions (CDRs) from aheavy chain variable region comprising an amino acid sequence shown inTables 1-4, or encoded by the nucleotide sequence in Tables 1-4. In oneembodiment, one or more of the CDRs (or collectively all of the CDRs)have one, two, three, four, five or more changes, e.g., amino acidsubstitutions, insertions, or deletions, relative to the amino acidsequence shown in Tables 1-4, or encoded by a nucleotide sequence shownin Tables 1-4. In certain embodiments, the anti-TIM-3 antibody moleculeincludes a substitution in a heavy chain CDR, e.g., one or moresubstitutions in a CDR1, CDR2 and/or CDR3 of the heavy chain. In oneembodiment, the anti-TIM-3 antibody molecule includes a substitution inthe heavy chain CDR2 at position 55 of the heavy chain region, e.g., asubstitution of an asparagine to serine, or an asparagine to glutamine,at position 55 of the heavy chain region according to Tables 1-4 (e.g.,any of SEQ ID NOs: 1 or 18 for murine or humanized, unmodified; or anyof SEQ ID NOs: 26, or 32 for a modified sequence).

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three complementarity determining regions (CDRs) from alight chain variable region of an antibody described herein, e.g., anantibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4, or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, or three CDRs (or collectively all of the CDRs) from alight chain variable region comprising an amino acid sequence shown inTables 1-4, or encoded by a nucleotide sequence shown in Tables 1-4. Insome embodiments, one or more of the CDRs (or collectively all of theCDRs) have one, two, three, four, five, six or more changes, e.g., aminoacid substitutions, insertions, or deletions, relative to the CDRs shownin Tables 1-4, or encoded by a nucleotide sequence shown in Tables 1-4.In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three CDRs (or collectively all of the CDRs) from a lightchain variable region comprising an amino acid sequence shown in Tables1-4, or encoded by a nucleotide sequence shown in Tables 1-4. In someembodiments, one or more of the CDRs (or collectively all of the CDRs)have one, two, three, four, five, six or more changes, e.g., amino acidsubstitutions, insertions, or deletions, relative to the CDRs shown inTables 1-4, or encoded by a nucleotide sequence shown in Tables 1-4.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, three, four, five or six CDRs (or collectively all of theCDRs) from a heavy and light chain variable region comprising an aminoacid sequence shown in Tables 1-4, or encoded by a nucleotide sequenceshown in Tables 1-4. In some embodiments, one or more of the CDRs (orcollectively all of the CDRs) have one, two, three, four, five, six ormore changes, e.g., amino acid substitutions, insertions, or deletions,relative to the CDRs shown in Tables 1-4, or encoded by a nucleotidesequence shown in Tables 1-4.

In certain embodiments, the anti-TIM-3 antibody molecule includes allsix CDRs from an antibody described herein, e.g., an antibody chosenfrom any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; oras described in Tables 1-4; or encoded by the nucleotide sequence inTables 1-4, or closely related CDRs, e.g., CDRs which are identical orwhich have at least one amino acid alteration, but not more than two,three or four alterations (e.g., substitutions, deletions, orinsertions, e.g., conservative substitutions). In certain embodiments,the anti-TIM-3 antibody molecule may include any CDR described herein.In certain embodiments, the anti-TIM-3 antibody molecule includes asubstitution in a heavy chain CDR, e.g., one or more substitutions in aCDR1, CDR2 and/or CDR3 of the heavy chain. In one embodiment, theanti-TIM-3 antibody molecule includes a substitution in the heavy chainCDR2 at position 55 of the heavy chain region, e.g., a substitution ofan asparagine to serine, or an asparagine to glutamine, at position 55of the heavy chain region according to Tables 1-4 (e.g., any of SEQ IDNOs: 1 or 18 for murine or humanized, unmodified; or any of SEQ ID NOs:26, or 32 for a modified sequence).

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three CDRs according to Kabat et al. (e.g., at least one,two, or three CDRs according to the Kabat definition as set out inTables 1-4) from a heavy chain variable region of an antibody describedherein, e.g., an antibody chosen from any of ABTIM3, ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4; or encoded bythe nucleotide sequence in Tables 1-4; or a sequence substantiallyidentical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% orhigher identical) to any of the aforesaid sequences; or which have atleast one amino acid alteration, but not more than two, three or fouralterations (e.g., substitutions, deletions, or insertions, e.g.,conservative substitutions) relative to one, two, or three CDRsaccording to Kabat et al. shown in Tables 1-4.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, or three CDRs according to Kabat et al. (e.g., at leastone, two, or three CDRs according to the Kabat definition as set out inTables 1-4) from a light chain variable region of an antibody describedherein, e.g., an antibody chosen from any of ABTIM, ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4; or encoded bythe nucleotide sequence in Tables 1-4; or a sequence substantiallyidentical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% orhigher identical) to any of the aforesaid sequences; or which have atleast one amino acid alteration, but not more than two, three or fouralterations (e.g., substitutions, deletions, or insertions, e.g.,conservative substitutions) relative to one, two, or three CDRsaccording to Kabat et al. shown in Tables 1-4.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, three, four, five, or six CDRs according to Kabat et al.(e.g., at least one, two, three, four, five, or six CDRs according tothe Kabat definition as set out in Tables 1-4) from the heavy and lightchain variable regions of an antibody described herein, e.g., anantibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4; or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences; or which have at least oneamino acid alteration, but not more than two, three or four alterations(e.g., substitutions, deletions, or insertions, e.g., conservativesubstitutions) relative to one, two, three, four, five, or six CDRsaccording to Kabat et al. shown in Tables 1-4.

In some embodiments, the anti-TIM-3 antibody molecule includes all sixCDRs according to Kabat et al. (e.g., all six CDRs according to theKabat definition as set out in Tables 1-4) from the heavy and lightchain variable regions of an antibody described herein, e.g., anantibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4; or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences; or which have at least oneamino acid alteration, but not more than two, three or four alterations(e.g., substitutions, deletions, or insertions, e.g., conservativesubstitutions) relative to all six CDRs according to Kabat et al. shownin Tables 1-4. In one embodiment, the anti-TIM-3 antibody molecule mayinclude any CDR described herein.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three Chothia hypervariable loops (e.g., at least one, two,or three hypervariable loops according to the Chothia definition as setout in Tables 1-4) from a heavy chain variable region of an antibodydescribed herein, e.g., an antibody chosen from any of ABTIM3,ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05,ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10,ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15,ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20,ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4;or encoded by the nucleotide sequence in Tables 1-4; or at least theamino acids from those hypervariable loops that contact TIM-3; or whichhave at least one amino acid alteration, but not more than two, three orfour alterations (e.g., substitutions, deletions, or insertions, e.g.,conservative substitutions) relative to one, two, or three hypervariableloops according to Chothia et al. shown in Tables 1-4.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, or three Chothia hypervariable loops (e.g., at leastone, two, or three hypervariable loops according to the Chothiadefinition as set out in Tables 1-4) of a light chain variable region ofan antibody described herein, e.g., an antibody chosen from any ofABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04,ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09,ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14,ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19,ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as describedin Tables 1-4; or encoded by the nucleotide sequence in Tables 1-4; orat least the amino acids from those hypervariable loops that contactTIM-3; or which have at least one amino acid alteration, but not morethan two, three or four alterations (e.g., substitutions, deletions, orinsertions, e.g., conservative substitutions) relative to one, two, orthree hypervariable loops according to Chothia et al. shown in Tables1-4.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, three, four, five, or six hypervariable loops (e.g., atleast one, two, three, four, five, or six hypervariable loops accordingto the Chothia definition as set out in Tables 1-4) from the heavy andlight chain variable regions of an antibody described herein, e.g., anantibody chosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02,ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07,ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12,ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17,ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22,ABTIM3-hum23; or as described in Tables 1-4; or encoded by thenucleotide sequence in Tables 1-4; or at least the amino acids fromthose hypervariable loops that contact TIM-3; or which have at least oneamino acid alteration, but not more than two, three or four alterations(e.g., substitutions, deletions, or insertions, e.g., conservativesubstitutions) relative to one, two, three, four, five or sixhypervariable loops according to Chothia et al. shown in Tables 1-4.

In some embodiments, the anti-TIM-3 antibody molecule includes all sixhypervariable loops (e.g., all six hypervariable loops according to theChothia definition as set out in Tables 1-4) of an antibody describedherein, e.g., an antibody chosen from any of ABTIM3, ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23; or closely related hypervariable loops,e.g., hypervariable loops which are identical or which have at least oneamino acid alteration, but not more than two, three or four alterations(e.g., substitutions, deletions, or insertions, e.g., conservativesubstitutions); or which have at least one amino acid alteration, butnot more than two, three or four alterations (e.g., substitutions,deletions, or insertions, e.g., conservative substitutions) relative toall six hypervariable loops according to Chothia et al. shown in Tables1-4. In one embodiment, the anti-TIM-3 antibody molecule may include anyhypervariable loop described herein.

In still another embodiment, the anti-TIM-3 antibody molecule includesat least one, two, or three hypervariable loops that have the samecanonical structures as the corresponding hypervariable loop of anantibody described herein, e.g., an antibody chosen from any of ABTIM3,ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05,ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10,ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15,ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20,ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, e.g., the same canonicalstructures as at least loop 1 and/or loop 2 of the heavy and/or lightchain variable domains of an antibody described herein. See, e.g.,Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinson et al.,(1992) J. Mol. Biol. 227:776-798 for descriptions of hypervariable loopcanonical structures. These structures can be determined by inspectionof the tables described in these references.

In certain embodiments, the anti-TIM-3 antibody molecule includes acombination of CDRs or hypervariable loops defined according to theKabat et al. and Chothia et al.

In one embodiment, the anti-TIM-3 antibody molecule includes at leastone, two or three CDRs or hypervariable loops from a heavy chainvariable region of an antibody described herein, e.g., an antibodychosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23,according to the Kabat and Chothia definition (e.g., at least one, two,or three CDRs or hypervariable loops according to the Kabat and Chothiadefinition as set out in Tables 1-4); or encoded by the nucleotidesequence in Tables 1-4; or a sequence substantially identical (e.g., atleast 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to anyof the aforesaid sequences; or which have at least one amino acidalteration, but not more than two, three or four alterations (e.g.,substitutions, deletions, or insertions, e.g., conservativesubstitutions) relative to one, two, or three CDRs or hypervariableloops according to Kabat and/or Chothia shown in Tables 1-4.

In nother embodiment, the anti-TIM-3 antibody molecule includes at leastone, two or three CDRs or hypervariable loops from a light chainvariable region of an antibody described herein, e.g., an antibodychosen from any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23,according to the Kabat and Chothia definition (e.g., at least one, two,or three CDRs or hypervariable loops according to the Kabat and Chothiadefinition as set out in Tables 1-4); or encoded by the nucleotidesequence in Tables 1-4; or a sequence substantially identical (e.g., atleast 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to anyof the aforesaid sequences; or which have at least one amino acidalteration, but not more than two, three or four alterations (e.g.,substitutions, deletions, or insertions, e.g., conservativesubstitutions) relative to one, two, or three CDRs or hypervariableloops according to Kabat and/or Chothia shown in Tables 1-4.

The anti-TIM-3 antibody molecule can contain any combination of CDRs orhypervariable loops according to the Kabat and Chothia definitions.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three Chothia hypervariable loops from a heavy chainvariable region of an antibody described herein, e.g., an antibody ofTables 1-4, or at least the amino acids from those hypervariable loopsthat contact TIM-3.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three Chothia hypervariable loops from a light chainvariable region of an antibody described herein, e.g., an antibody ofTables 1-4, or at least the amino acids from those hypervariable loopsthat contact TIM-3.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three Kabat hypervariable loops from a heavy chain variableregion of an antibody described herein, e.g., an antibody of Tables 1-4,or at least the amino acids from those hypervariable loops that contactTIM-3.

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three Kabat hypervariable loops from a light chain variableregion of an antibody described herein, e.g., an antibody of Tables 1-4,or at least the amino acids from those hypervariable loops that contactTIM-3.

In certain embodiments, the anti-TIM-3 antibody molecule includes atleast one, two, three, four, five, or six hypervariable loops from theheavy and light chain variable regions of an antibody described herein,e.g., an antibody of Tables 1-4, or at least the amino acids from thosehypervariable loops that contact TIM-3.

In certain embodiments, the anti-TIM-3 antibody molecule includes allsix hypervariable loops from the heavy and light chain variable regionsof an antibody described herein, e.g., an antibody of Tables 1-4, or atleast the amino acids from those hypervariable loops that contact TIM-3,or at least the amino acids from those hypervariable loops that contactTIM-3, or closely related hypervariable loops, e.g., hypervariable loopswhich are identical or which have at least one amino acid alteration,but not more than two, three or four alterations (e.g., substitutions,e.g., conservative substitutions, deletions, or insertions).

In some embodiments, the anti-TIM-3 antibody molecule includes at leastone, two, or three hypervariable loops that have the same canonicalstructures as the corresponding hypervariable loop of an antibodydescribed herein, e.g., an antibody of Tables 1-4, e.g., the samecanonical structures as at least loop 1 and/or loop 2 of the heavyand/or light chain variable domains of an antibody described herein.See, e.g., Chothia et al., (1992) J. Mol. Biol. 227:799-817; Tomlinsonet al., (1992) J. Mol. Biol. 227:776-798 for descriptions ofhypervariable loop canonical structures. These structures can bedetermined by inspection of the tables described in these references. Inan embodiment, e.g., an embodiment comprising a variable region, CDR(e.g., Chothia CDR or Kabat CDR), or other sequence referred to herein,e.g., in Tables 1-4, the antibody molecule is a monospecific antibodymolecule, a bispecific antibody molecule, or is an antibody moleculethat comprises an antigen binding fragment of an antibody, e.g., a halfantibody or antigen binding fragment of a half antibody. In certainembodiments the antibody molecule is a bispecific antibody moleculehaving a first binding specificity for TIM-3 and a second bindingspecificity for PD-1, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/orCEACAM-5), PD-L1 or PD-L2.

In certain embodiments, the light or the heavy chain variable framework(e.g., the region encompassing at least FR1, FR2, FR3, or FR4) of theanti-TIM-3 antibody molecule can be chosen from: (a) a light or heavychain variable framework including at least 80%, 85%, 87% 90%, 92%, 93%,95%, 97%, 98%, or preferably 100% of the amino acid residues from ahuman light or heavy chain variable framework, e.g., a light or heavychain variable framework residue from a human mature antibody, a humangermline sequence, or a human consensus sequence; (b) a light or heavychain variable framework including from 20% to 80%, 40% to 60%, 60% to90%, or 70% to 95% of the amino acid residues from a human light orheavy chain variable framework, e.g., a light or heavy chain variableframework residue from a human mature antibody, a human germlinesequence, or a human consensus sequence; (c) a non-human framework(e.g., a rodent framework); or (d) a non-human framework that has beenmodified, e.g., to remove antigenic or cytotoxic determinants, e.g.,deimmunized, or partially humanized. In some embodiments, the light orheavy chain variable framework region includes a light or heavy chainvariable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94,95, 96, 97, 98, 99% identical or identical to the frameworks of a VL orVH segment of a human germline gene.

In certain embodiments, the anti-TIM-3 antibody molecule comprises aheavy chain variable domain having at least one, two, three, four, five,six, seven, ten, fifteen, twenty or more changes, e.g., amino acidsubstitutions, insertions, or deletions, from an amino acid sequence of,e.g., the amino acid sequence of the FR region in the entire variableregion, e.g., shown in FIG. 1A. In some embodiments, the anti-TIM-3antibody molecule comprises a heavy chain variable domain having one ormore (e.g., all) of: A at position 2, Y at position 3, S at position 7,R at position 13, V at position 37, R at position 42, V at position 72,A at position 79, or F at position 95, e.g., the amino acid sequence ofthe FR in the entire variable region, e.g., as shown in FIG. 1A. In someembodiments, the anti-TIM-3 antibody molecule comprises a heavy chainvariable domain having 2, 3, 4, 5, 6, 7, 8, or 9 positions selectedfrom: A at position 2, Y at position 3, S at position 7, R at position13, V at position 37, R at position 42, V at position 72, A at position79, or F at position 95 of the amino acid sequence of an antibody ofTables 1-4, e.g.,

In certain embodiments (and optionally in combination with the heavychain substitutions described herein, e.g., in the previous paragraph),the anti-TM-3 antibody molecule comprises a light chain variable domainhaving at least one, two, three, four, five, six, seven, ten, fifteen,twenty or more amino acid changes, e.g., amino acid substitutions,insertions, or deletions, from an amino acid sequence of Tables 1-4,e.g., the amino acid sequence of the FR region in the entire variableregion, e.g., shown in FIG. 1B. In certain embodiments, the anti-TIM-3antibody comprises a light chain variable domain having M at position 89of the amino acid sequence of an antibody of Tables 1-4.

In some embodiments, the heavy or light chain variable domain, or both,of the of the anti-TIM-3 antibody molecule includes an amino acidsequence, which is substantially identical to an amino acid disclosedherein, e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical to a variable region of an antibody described herein, e.g., anantibody chosen from any of ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; oras described in Tables 1-4; or encoded by the nucleotide sequence inTables 1-4; or which differs at least 1 or 5 residues, but less than 40,30, 20, or 10 residues, from a variable region of an antibody describedherein.

In certain embodiments, the heavy or light chain variable region, orboth, of the anti-TIM-3 antibody molecule includes an amino acidsequence encoded by a nucleic acid sequence described herein or anucleic acid that hybridizes to a nucleic acid sequence described herein(e.g., a nucleic acid sequence as shown in Tables 1-4) or itscomplement, e.g., under low stringency, medium stringency, or highstringency, or other hybridization condition described herein.

In certain embodiments, the anti-TIM-3 antibody molecule comprises atleast one, two, three, or four antigen-binding regions, e.g., variableregions, having an amino acid sequence as set forth in Tables 1-4, or asequence substantially identical thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, or which differs byno more than 1, 2, 5, 10, or 15 amino acid residues from the sequencesshown in Tables 1-4. In certain embodiments, the anti-TIM-3 antibodymolecule includes a VH and/or VL domain encoded by a nucleic acid havinga nucleotide sequence that encodes an antibody of Tables 1-4, or asequence substantially identical to any one of the nucleotide sequences(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, or which differs by no more than 3, 6, 15, 30, or 45nucleotides from the sequences shown in Tables 1-4).

In certain embodiments, the anti-TIM-3 antibody molecule comprises atleast one, two, or three (e.g., all) CDRs from a heavy chain variableregion having an amino acid sequence as set forth in Tables 1-4, or asequence substantially homologous thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, and/or having one,two, three or more substitutions, insertions or deletions, e.g.,conserved substitutions). In some embodiments, the anti-TIM-3 antibodymolecule comprises at least one, two, or three (e.g., all) CDRs from alight chain variable region having an amino acid sequence as set forthin Tables 1-4, or a sequence substantially homologous thereto (e.g., asequence at least about 85%, 90%, 95%, 99% or more identical thereto,and/or having one, two, three or more substitutions, insertions ordeletions, e.g., conserved substitutions). In certain embodiments, theanti-TIM-3 antibody molecule comprises at least one, two, three, four,five or six (e.g., all) CDRs from heavy and light chain variable regionshaving an amino acid sequence as set forth in Tables 1-4, or a sequencesubstantially homologous thereto (e.g., a sequence at least about 85%,90%, 95%, 99% or more identical thereto, and/or having one, two, threeor more substitutions, insertions or deletions, e.g., conservedsubstitutions).

In some embodiments, the anti-TIM-3 antibody molecule comprises at leastone, two, or three (e.g., all) CDRs and/or hypervariable loops from aheavy chain variable region having an amino acid sequence of an antibodydescribed herein, e.g., an antibody chosen from any of ABTIM3-hum01,ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06,ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11,ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16,ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21,ABTIM3-hum22, ABTIM3-hum23, as summarized in Tables 1-4, or a sequencesubstantially identical thereto (e.g., a sequence at least about 85%,90%, 95%, 99% or more identical thereto, and/or having one, two, threeor more substitutions, insertions or deletions, e.g., conservedsubstitutions). In certain embodiments, the anti-TIM-3 antibody moleculecomprises at least one, two, or three (e.g., all) CDRs and/orhypervariable loops from a light chain variable region having an aminoacid sequence of an antibody described herein, e.g., an antibody chosenfrom any of ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04,ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09,ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14,ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19,ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as summarized inTables 1-4, or a sequence substantially identical thereto (e.g., asequence at least about 85%, 90%, 95%, 99% or more identical thereto,and/or having one, two, three or more substitutions, insertions ordeletions, e.g., conserved substitutions). In some embodiments, theanti-TIM-3 antibody molecule comprises all six CDRs and/or hypervariableloops described herein, e.g., described in Tables 1-4.

In some embodiments, the antibody molecule has a variable region that isidentical in sequence, or which differs by 1, 2, 3, or 4 amino acidsfrom a variable region described herein (e.g., an FR region disclosedherein).

In some embodiments, the anti-TIM-3 antibody molecule is a full antibodyor fragment thereof (e.g., a Fab, F(ab′)2, Fv, or a single chain Fvfragment (scFv)). In certain embodiments, the anti-TIM-3 antibodymolecule is a monoclonal antibody or an antibody with singlespecificity. The anti-TIM-3 antibody molecule can also be a humanized,chimeric, camelid, shark, or in vitro-generated antibody molecule. Insome embodiments, the anti-TIM-3 antibody molecule thereof is ahumanized antibody molecule. The heavy and light chains of theanti-TIM-3 antibody molecule can be full-length (e.g., an antibody caninclude at least one or at least two complete heavy chains, and at leastone or at least two complete light chains) or can include anantigen-binding fragment (e.g., a Fab, F(ab′)2, Fv, a single chain Fvfragment, a single domain antibody, a diabody (dAb), a bivalent orbispecific antibody or fragment thereof, a single domain variantthereof, or a camelid antibody).

In certain embodiments, the anti-TIM-3 antibody molecule is in the formof a bispecific or multispecific antibody molecule. In one embodiment,the bispecific antibody molecule has a first binding specificity toTIM-3 and a second binding specifity, e.g., a second binding specificityto PD-1, LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), PD-L1 or PD-L2.In one embodiment, the bispecific antibody molecule binds to TIM-3 andPD-1. In another embodiment, the bispecific antibody molecule binds toTIM-3 and LAG-3. In another embodiment, the bispecific antibody moleculebinds to TIM-3 and CEACAM (e.g., CEACAM-1,-3 and/or -5). In anotherembodiment, the bispecific antibody molecule binds to TIM-3 andCEACAM-1. In another embodiment, the bispecific antibody molecule bindsto TIM-3 and CEACAM-3. In yet another embodiment, the bispecificantibody molecule binds to TIM-3 and CEACAM-5. In another embodiment,the bispecific antibody molecule binds to TIM-3 and PD-L1. In yetanother embodiment, the bispecific antibody molecule binds to TIM-3 andPD-L2. Any combination of the aforesaid molecules can be made in amultispecific antibody molecule, e.g., a trispecific antibody thatincludes a first binding specificity to TIM-3, and a second and thirdbinding specifities to one or more of: PD-1, LAG-3, CEACAM (e.g.,CEACAM-1, -3 and/or -5), PD-L1 or PD-L2.

In other embodiments, the anti-TIM-3 antibody molecule is used incombination with a bispecific molecule comprising one or more of: PD-1,LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), PD-L1 or PD-L2. In oneembodiment, the bispecific antibody molecule used in combination bindsto CEACAM (e.g., CEACAM-1, -3 and/or -5) and LAG-3. In anotherembodiment, the bispecific antibody molecule used in combination bindsto CEACAM (e.g., CEACAM-1, -3 and/or -5) and PD-1. In anotherembodiment, the bispecific antibody molecule used in combination bindsto LAG-3 and PD-1.

In certain embodiments, the anti-TIM-3 antibody molecule has a heavychain constant region (Fc) chosen from, e.g., the heavy chain constantregions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE;particularly, chosen from, e.g., the heavy chain constant regions ofIgG1, IgG2, IgG3, and IgG4, more particularly, the heavy chain constantregion of IgG1 or IgG2 (e.g., human IgG1 or IgG2). In some embodiments,the heavy chain constant region is human IgG1. In some embodiments, theanti-TIM-3 antibody molecule has a light chain constant region chosenfrom, e.g., the light chain constant regions of kappa or lambda, in someembodiments kappa (e.g., human kappa). In some embodiments, the constantregion is altered, e.g., mutated, to modify the properties of theanti-TIM-3 antibody molecule (e.g., to increase or decrease one or moreof: Fc receptor binding, antibody glycosylation, the number of cysteineresidues, effector cell function, or complement function). For example,the constant region may be mutated at positions 296 (M to Y), 298 (S toT), 300 (T to E), 477 (H to K) and 478 (N to F) to alter Fc receptorbinding (e.g., the mutated positions correspond to positions 132 (M toY), 134 (S to T), 136 (T to E), 313 (H to K) and 314 (N to F) of SEQ IDNOs: 108 or 110; or positions 135 (M to Y), 137 (S to T), 139 (T to E),316 (H to K) and 317 (N to F) of SEQ ID NOs: 111, 112, 113 or 114). Inanother embodiment, the heavy chain constant region of an IgG4, e.g., ahuman IgG4, is mutated at position 228 according to EU numbering (e.g.,S to P), e.g., as shown in Table 5. In certain embodiments, theanti-TIM-3 antibody molecules comprises a human IgG4 mutated at position228 according to EU numbering (e.g., S to P), e.g., as shown in Table 5;and a kappa light chain constant region, e.g., as shown in Table 5. Instill another embodiment, the heavy chain constant region of an IgG1,e.g., a human IgG1, is mutated at one or more of position 297 (e.g., Nto A), position 265 (e.g., D to A), position 329 (e.g., P to A),position 234 (e.g., L to A), or position 235 (e.g., L to A), allaccording to EU numbering, e.g., as shown in Table 5. In certainembodiments, the anti-TIM-3 antibody molecules comprises a human IgG1mutated at one or more of the aforesaid positions, e.g., as shown inTable 5; and a kappa light chain constant region, e.g., as shown inTable 5. In some embodiments, the anti-TIM-3 antibody molecule is ahumanized antibody molecule.

In some embodiments, the anti-TIM-3 antibody molecules comprisecombinations of human or humanized framework regions with CDRs(complementarity determining regions).

The invention also features an antibody molecule that competes with amonoclonal antibody, e.g., an antibody molecule described herein, forbinding to human TIM-3.

In certain embodiments, the monoclonal antibody comprises:

(a) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQID NO: 10; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and a lightchain variable region (VL) comprising a VLCDR1 amino acid sequence ofSEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and aVLCDR3 amino acid sequence of SEQ ID NO: 14;

(b) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 4; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(c) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 25; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14;

(d) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 24; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(e) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 31; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14; or

(f) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 30; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8.

The invention also features an antibody molecule that binds to the same(or substantially the same) or an overlapping (or substantiallyoverlapping) epitope as a monoclonal antibody, e.g., an antibodymolecule described herein, to human TIM-3.

In certain embodiments, the monoclonal antibody comprises:

(a) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQID NO: 10; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and a lightchain variable region (VL) comprising a VLCDR1 amino acid sequence ofSEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and aVLCDR3 amino acid sequence of SEQ ID NO: 14;

(b) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 4; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(c) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 25; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14;

(d) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 24; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(e) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 31; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14; or

(f) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 30; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8.

The invention also features a nucleic acid molecule that comprise one orboth nucleotide sequences that encode heavy and light chain variableregions, CDRs, hypervariable loops, framework regions of the anti-TIM-3antibody molecules, as described herein. In certain embodiments, thenucleotide sequence that encodes the anti-TIM-3 antibody molecule iscodon optimized. For example, the invention features a first and secondnucleic acid encoding heavy and light chain variable regions,respectively, of an anti-TIM-3 antibody molecule chosen from one or moreof, e.g., any of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, assummarized in Tables 1-4, or a sequence substantially identical thereto.For example, the nucleic acid can comprise a nucleotide sequence as setforth in Tables 1-4, or a sequence substantially identical thereto(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, or which differs by no more than 3, 6, 15, 30, or 45nucleotides from the sequences shown in Tables 1-4).

In some embodiments, nucleic acids comprising nucleotide sequences thatencode heavy and light chain variable regions and CDRs of the anti-TIM-3antibody molecules, as described herein, are disclosed. For example, thedisclosure provides a first and second nucleic acid encoding heavy andlight chain variable regions, respectively, of an anti-TIM-3 antibodymolecule according to Tables 1-4 or a sequence substantially identicalthereto. For example, the nucleic acid can comprise a nucleotidesequence encoding an an anti-TIM-3 antibody molecule according to Table1-4, or a sequence substantially identical to that nucleotide sequence(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, or which differs by no more than 3, 6, 15, 30, or 45nucleotides from the aforementioned nucleotide sequence.

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs, or hypervariableloops, from a heavy chain variable region having an amino acid sequenceas set forth in Tables 1-4, or a sequence substantially homologousthereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or moreidentical thereto, and/or having one, two, three or more substitutions,insertions or deletions, e.g., conserved substitutions).

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs, or hypervariableloops, from a light chain variable region having an amino acid sequenceas set forth in Tables 1-4, or a sequence substantially homologousthereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or moreidentical thereto, and/or having one, two, three or more substitutions,insertions or deletions, e.g., conserved substitutions).

In some embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, three, four, five, or six CDRs, orhypervariable loops, from heavy and light chain variable regions havingan amino acid sequence as set forth in Table 1-4, or a sequencesubstantially homologous thereto (e.g., a sequence at least about 85%,90%, 95%, 99% or more identical thereto, and/or having one, two, threeor more substitutions, insertions or deletions, e.g., conservedsubstitutions).

In some embodiments, the anti-TIM-3 antibody molecule is isolated orrecombinant.

In certain aspects, this disclosure features host cells and vectorscontaining the nucleic acids described herein. The nucleic acids may bepresent in a single vector or separate vectors present in the same hostcell or separate host cell. The host cell can be a eukaryotic cell,e.g., a mammalian cell, an insect cell, a yeast cell, or a prokaryoticcell, e.g., E. coli. For example, the mammalian cell can be a culturedcell or a cell line. Exemplary mammalian cells include lymphocytic celllines (e.g., NSO), Chinese hamster ovary cells (CHO), COS cells, oocytecells, and cells from a transgenic animal, e.g., mammary epithelialcell.

In some aspects, the present disclosure provides a method of providingan antibody molecule described herein. The method may include: providinga TIM-3 antigen (e.g., an antigen comprising at least a portion of aTIM-3 epitope, e.g., the IgV domain of TIM-3); obtaining an antibodymolecule that specifically binds to the TIM-3 antigen; and evaluating ifthe antibody molecule specifically binds to the TIM-3 antigen, orevaluating efficacy of the antibody molecule in modulating, e.g.,stimulating or inhibiting, the activity of TIM-3. The method can furtherinclude administering the antibody molecule to a subject, e.g., a humanor non-human animal.

In certain aspects, the disclosure provides, compositions, e.g.,pharmaceutical compositions, which include a pharmaceutically acceptablecarrier, excipient or stabilizer, and at least one of the anti-TIM-3antibody molecules described herein. In one embodiment, the composition,e.g., the pharmaceutical composition, includes a combination of theantibody molecule and one or more agents, e.g., a therapeutic agent orother antibody molecule, as described herein. In some embodiments, theantibody molecule is conjugated to a label or a therapeutic agent. Insome embodiments, the compositions, e.g., the pharmaceuticalcompositions, comprise a combination of the antibody molecule and asecond agent, e.g., a therapeutic agent, or two or more of the aforesaidantibody molecules, as further described herein.

The anti-TIM-3 antibody molecules disclosed herein can inhibit, reduceor neutralize one or more activities of TIM-3, e.g., resulting inblockade or reduction of an immune checkpoint on T cells or NK cells, orreinvigoration of an immune response by modulating antigen-presentingcells. In one embodiment, the antibody molecule results in one or moreof: enhancing IFN-gamma and/or TNF alpha section in T cells; enhancingproliferation in T cells, e.g., CD4+ or CD8+ T cells; enhancing NK cellcytotoxic activity; or reducing suppressor activity of regulatory Tcells (Tregs) or macrophages; or increasing capacity of macrophages ordendritic cells to stimulate an immune response. Thus, such antibodymolecules can be used to treat or prevent disorders where enhancing animmune response in a subject is desired.

Uses of the Anti-TIM-3 Antibody Molecules

The antibody molecules disclosed herein can modulate (e.g., enhance,stimulate, increase, inhibit, reduce or neutralize) one or moreactivities of TIM-3. In some embodiments, the antibody molecule resultsin one or more of: enhancing IFN-gamma secretion and/or proliferation inT cells or enhancing NK cell cytotoxic activity. For instance, in someembodiments, the anti-TIM-3 antibody molecule increases IFN-gammasecretion by at least 16%, 18%, 20%, 22%, 24%, 26%, 28%, or 30%, e.g.,in an assay of Example 4. In certain embodiments, the anti-TIM-3antibody molecule increases NK cell cytotoxic activity by at least about10%, 20%, 30%, 40%, 60%, 80%, or 100%, e.g., in an assay of Example 5.For example, the anti-TIM-3 antibody molecule could increase NK cellcytotoxic activity to at least about 60% or 70% of target cells killedwhen E/T=5, to at least about 75% or 85% of target cells killed whenE/T=12, or to at least about 85% or 95% of target cells killed whenE/T=25, e.g., in an assay of Example 5.

In certain aspects, a method of modulating (e.g., stimulating orinhibiting) an immune response in a subject is provided. The methodcomprises administering to the subject an anti-TIM-3 antibody moleculedisclosed herein, (e.g., a therapeutically effective amount of ananti-TIM-3 antibody molecule), alone or in combination with one or moreagents or procedures (e.g., in combination with other immunomodulatoryagents), such that the immune response in the subject is modulated. Insome embodiments, the antibody molecule enhances, stimulates orincreases an immune response in the subject. In some embodiments, theantibody molecule inhibits, reduces, or neutralizes an immune responsein a subject.

The subject can be a mammal, e.g., a monkey, a primate, preferably ahigher primate, e.g., a human (e.g., a patient having, or at risk ofhaving, a disorder described herein). In some embodiments, the subjectis in need of enhancing an immune response, and in some embodiments, thesubject is in need of inhibiting an immune response. In one embodiment,the subject has, or is at risk of, having a disorder described herein,e.g., a cancer or an infectious disorder as described herein. In certainembodiments, the subject is, or is at risk of being, immunocompromised.For example, the subject is undergoing or has undergone achemotherapeutic treatment and/or radiation therapy. Alternatively, orin combination, the subject is, or is at risk of being,immunocompromised as a result of an infection.

In one aspect, a method of treating (e.g., one or more of reducing,inhibiting, or delaying progression) a cancer or a tumor in a subject isprovided. The method comprises administering to the subject ananti-TIM-3 antibody molecule described herein, e.g., a therapeuticallyeffective amount of an anti-TIM-3 antibody molecule, alone or incombination with one or more agents or procedures. In certainembodiments, the anti-TIM-3 antibody molecule is administered incombination with a modulator of a costimulatory molecule (e.g., anagonist of a costimulatory molecule) or a modulator of an inhibitorymolecule (e.g., an inhibitor of an immune checkpoint inhibitor), e.g.,as described herein.

This disclosure also provides a method of reducing or inhibiting growthof a cancer or tumor cells (e.g., treating a cancer) in a subject,comprising administering to the subject an anti-TIM-3 antibody moleculedescribed herein, e.g., a therapeutically effective amount of ananti-TIM-3 antibody molecule, alone or in combination with a secondagent, e.g., an immunomodulator (e.g., an anti-PD-1, PD-L1, LAG-3 orCEACAM-1 inhibitor (e.g., antibody), or a combination thereof.

In certain embodiments, the cancer treated with the anti-TIM-3 antibodymolecule, alone or in combination with one or more immunomodulatos,includes but is not limited to, a solid tumor, a hematological cancer(e.g., leukemia, lymphoma, myeloma, e.g., multiple myeloma), and ametastatic lesion. In one embodiment, the cancer is a solid tumor.Examples of solid tumors include malignancies, e.g., sarcomas andcarcinomas, e.g., adenocarcinomas of the various organ systems, such asthose affecting the lung, breast, ovarian, lymphoid, gastrointestinal(e.g., colon), anal, genitals and genitourinary tract (e.g., renal,urothelial, bladder cells, prostate), pharynx, CNS (e.g., brain, neuralor glial cells), head and neck, skin (e.g., melanoma), and pancreas, aswell as adenocarcinomas which include malignancies such as coloncancers, rectal cancer, renal-cell carcinoma, liver cancer, non-smallcell lung cancer, cancer of the small intestine and cancer of theesophagus. The cancer may be at an early, intermediate, late stage ormetastatic cancer.

In one embodiment, the cancer is chosen from a lung cancer (e.g., lungadenocarcinoma or a non-small cell lung cancer (NSCLC) (e.g., a NSCLCwith squamous and/or non-squamous histology, or a NSCLCadenocarcinoma)), a melanoma (e.g., an advanced melanoma), a renalcancer (e.g., a renal cell carcinoma), a liver cancer (e.g.,hepatocellular carcinoma), a myeloma (e.g., a multiple myeloma), aprostate cancer, a breast cancer (e.g., a breast cancer that does notexpress one, two or all of estrogen receptor, progesterone receptor, orHer2/neu, e.g., a triple negative breast cancer), an ovarian cancer, acolorectal cancer, a pancreatic cancer, a head and neck cancer (e.g.,head and neck squamous cell carcinoma (HNSCC), anal cancer,gastro-esophageal cancer (e.g., esophageal squamous cell carcinoma),mesothelioma, nasopharyngeal cancer, thyroid cancer, cervical cancer, alymphoproliferative disease (e.g., a post-transplant lymphoproliferativedisease) or a hematological cancer, (e.g., diffuse large B celllymphoma, T-cell lymphoma, B-cell lymphoma, or a non-Hogdkin lymphoma),or a leukemia (e.g., a myeloid leukemia or a lymphoid leukemia).

In another embodiment, the cancer is chosen form a carcinoma (e.g.,advanced or metastatic carcinoma), melanoma or a lung carcinoma, e.g., anon-small cell lung carcinoma.

In one embodiment, the cancer is a lung cancer, e.g., a lungadenocarcinoma, non-small cell lung cancer or small cell lung cancer.

In one embodiment, the cancer is a melanoma, e.g., an advanced melanoma.In one embodiment, the cancer is an advanced or unresectable melanomathat does not respond to other therapies. In other embodiments, thecancer is a melanoma with a BRAF mutation (e.g., a BRAF V600 mutation).In yet other embodiments, the anti-TIM-3 antibody molecule isadministered after treatment with an anti-CTLA-4 antibody (e.g.,ipilimumab) with or without a BRAF inhibitor (e.g., vemurafenib ordabrafenib).

In another embodiment, the cancer is a hepatocarcinoma, e.g., anadvanced hepatocarcinoma, with or without a viral infection, e.g., achronic viral hepatitis.

In another embodiment, the cancer is a prostate cancer, e.g., anadvanced prostate cancer.

In yet another embodiment, the cancer is a myeloma, e.g., multiplemyeloma.

In yet another embodiment, the cancer is a renal cancer, e.g., a renalcell carcinoma (RCC) (e.g., a metastatic RCC, clear cell renal cellcarcinoma (CCRCC) or kidney papillary cell carcinoma).

In one embodiment, the cancer microenvironment has an elevated level ofPD-L1 expression. Alternatively, or in combination, the cancermicroenvironment can have increased IFNγ and/or CD8 expression.

In some embodiments, the subject has, or is identified as having, atumor that has one or more of high PD-L1 level or expression, or asbeing Tumor Infiltrating Lymphocyte (TIL)+ (e.g., as having an increasednumber of TILs), or both. In certain embodiments, the subject has, or isidentified as having, a tumor that has high PD-L1 level or expressionand that is TIL+. In some embodiments, the methods described hereinfurther include identifying a subject based on having a tumor that hasone or more of high PD-L1 level or expression or as being TIL+, or both.In certain embodiments, the methods described herein further includeidentifying a subject based on having a tumor that has high PD-L1 levelor expression and as being TIL+. In some embodiments, tumors that areTIL+ are positive for CD8 and IFNγ. In some embodiments, the subjecthas, or is identified as having, a high percentage of cells that arepositive for one, two or more of PD-L1, CD8, and/or IFNγ. In certainembodiments, the subject has or is identified as having a highpercentage of cells that are positive for all of PD-L1, CD8, and IFNγ.

In some embodiments, the methods described herein further includeidentifying a subject based on having a high percentage of cells thatare positive for one, two or more of PD-L1, CD8, and/or IFNγ. In certainembodiments, the methods described herein further include identifying asubject based on having a high percentage of cells that are positive forall of PD-L1, CD8, and IFNγ. In some embodiments, the subject has, or isidentified as having, one, two or more of PD-L1, CD8, and/or IFNγ, andone or more of a lung cancer, e.g., squamous cell lung cancer or lungadenocarcinoma; a head and neck cancer; a squamous cell cervical cancer;a stomach cancer; an esophageal cancer; a thyroid cancer; a melanoma,and/or a nasopharyngeal cancer (NPC). In certain embodiments, themethods described herein further describe identifying a subject based onhaving one, two or more of PD-L1, CD8, and/or IFNγ, and one or more of alung cancer, e.g., squamous cell lung cancer or lung adenocarcinoma; ahead and neck cancer; a squamous cell cervical cancer; a stomach cancer;a thyroid cancer; a melanoma, and/or a nasopharyngeal cancer.

In some embodiments, subject has, or is identified as having, a tumorthat has one, two, or more of high PD-1 level or expression, high TIM-3level or expression, and/or high level of infiltration of regulatory Tcells in the tumor, e.g., an increased number or percentage of Tregspresent in the tumor. In certain embodiments, the subject has, or isidentified as having, a tumor that has a high level or expression ofPD-1 and TIM-3, and a high level, e.g., number, or regulatory T cells inthe tumor. In some embodiments, the methods described herein furtherinclude identifying a subject based on one, two or more of a highpercentage of cells that are positive for PD-1, a high percentage ofcells that are positive for TIM-3, and/or a high level of infiltrationof regulatory T cells in the tumor, e.g., an increased number orpercentage of Tregs present in the tumor. In some embodiments, themethods described herein further include identifying a subject based onone, two or more of a high percentage of cells that are positive forPD-1, a high percentage of cells that are positive for TIM-3, and/or ahigh level of infiltration of regulatory T cells in the tumor, e.g., anincreased number or percentage of Tregs present in the tumor, and one ormore of a lung cancer, e.g., non-small cell lung cancer (NSCLC); ahepatocellular cancer, e.g., hepatocellular carcinoma; or an ovariancancer, e.g., ovarian carcinoma.

Methods and compositions disclosed herein are useful for treatingmetastatic lesions associated with the aforementioned cancers.

In further aspects, this disclosure provides a method of treating aninfectious disease in a subject, comprising administering to a subject atherapeutically effective amount of an anti-TIM-3 antibody describedherein, or antigen-binding portion thereof, alone or in combination withone or more agents or procedures (e.g., one or more immunomodulatoryagents).

Still further, this disclosure provides methods of enhancing an immuneresponse to an antigen in a subject, comprising administering to thesubject: (i) the antigen; and (ii) an anti-TIM-3 antibody, orantigen-binding portion thereof, such that an immune response to theantigen in the subject is enhanced. The antigen can be, for example, atumor antigen, a viral antigen, a bacterial antigen or an antigen from apathogen.

The anti-TIM-3 antibody molecule can be administered to the subjectsystemically (e.g., orally, parenterally, subcutaneously, intravenously,rectally, intramuscularly, intraperitoneally, intranasally,transdermally, or by inhalation or intracavitary installation),topically, or by application to mucous membranes, such as the nose,throat and bronchial tubes.

The anti-TIM-3 antibody molecule can be used alone in unconjugated form,or can be bound to a substance, e.g., a cytotoxic agent or moiety (e.g.,a therapeutic drug; a compound emitting radiation; molecules of plant,fungal, or bacterial origin; or a biological protein (e.g., a proteintoxin) or particle (e.g., a recombinant viral particle, e.g., via aviral coat protein). For example, the anti-TIM-3 antibody can be coupledto a radioactive isotope such as an α-, β-, or γ-emitter, or a β- andγ-emitter.

Dosages and therapeutic regimens of the anti-TIM-3 antibody molecule canbe determined by a skilled artisan. In certain embodiments, theanti-TIM-3 antibody molecule is administered by injection (e.g.,subcutaneously or intravenously) at a dose of about 1 to 30 mg/kg, e.g.,about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about3 mg/kg. The dosing schedule can vary from e.g., once a week to onceevery 2, 3, or 4 weeks. In one embodiment, the anti-TIM-3 antibodymolecule is administered at a dose from about 10 to 20 mg/kg every otherweek.

The antibody molecules described herein are preferred for use in themethods described herein, although other anti-TIM-3 antibodies can beused instead, or in combination with an anti-TIM-3 antibody molecule ofthe invention.

Combination Therapies

The methods and compositions described herein can be used in combinationwith other therapeutic modalities. In some embodiments, the methods ofdescribed herein include administering to the subject an anti-TIM-3antibody molecule as described herein, in combination with a cytotoxicagent, in an amount effective to treat or prevent said disorder. Theantibody molecule and the cytotoxic agent can be administeredsimultaneously or sequentially.

Any combination and sequence of the anti-TIM-3 antibody molecules andother therapeutic modalities can be used. The anti-TIM-3 antibodymolecule and/or other therapeutic modalities can be administered duringperiods of active disorder, or during a period of remission or lessactive disease. The anti-TIM-3 antibody molecule and other therapeuticmodalities can be administered before treatment, concurrently withtreatment, post-treatment, or during remission of the disorder.

In certain embodiments, the methods and compositions described hereinare administered in combination with one or more of other antibodymolecules, chemotherapy, other anti-cancer therapy (e.g., targetedanti-cancer therapies, gene therapy, viral therapy, RNA therapy bonemarrow transplantation, nanotherapy, or oncolytic drugs), cytotoxicagents, immune-based therapies (e.g., cytokines or cell-based immunetherapies), surgical procedures (e.g., lumpectomy or mastectomy) orradiation procedures, or a combination of any of the foregoing. Theadditional therapy may be in the form of adjuvant or neoadjuvanttherapy. In some embodiments, the additional therapy is an enzymaticinhibitor (e.g., a small molecule enzymatic inhibitor) or a metastaticinhibitor. Exemplary cytotoxic agents that can be administered incombination with include antimicrotubule agents, topoisomeraseinhibitors, anti-metabolites, mitotic inhibitors, alkylating agents,anthracyclines, vinca alkaloids, intercalating agents, agents capable ofinterfering with a signal transduction pathway, agents that promoteapoptosis, proteosome inhibitors, and radiation (e.g., local or wholebody irradiation (e.g., gamma irradiation). In other embodiments, theadditional therapy is surgery or radiation, or a combination thereof. Inother embodiments, the additional therapy is a therapy targeting one ormore of PI3K/AKT/mTOR pathway, an HSP90 inhibitor, or a tubulininhibitor.

Alternatively, or in combination with the aforesaid combinations, themethods and compositions described herein can be administered incombination with one or more of: an immunomodulator (e.g., an activatorof a costimulatory molecule or an inhibitor of an inhibitory molecule,e.g., an immune checkpoint molecule); a vaccine, e.g., a therapeuticcancer vaccine; or other forms of cellular immunotherapy.

Exemplary non-limiting combinations and uses of the anti-TIM-3 antibodymolecules include the following.

In certain embodiments, the anti-TIM-3 antibody molecule is administeredin combination with a modulator of a costimulatory molecule or aninhibitory molecule, e.g., a co-inhibitory ligand or receptor.

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with a modulator, e.g., agonist, of a costimulatorymolecule. In one embodiment, the agonist of the costimulatory moleculeis chosen from an agonist (e.g., an agonistic antibody orantigen-binding fragment thereof, or a soluble fusion) of OX40, CD2,CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137),GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160,B7-H3 or CD83 ligand.

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with an inhibitor of an inhibitory (or immune checkpoint)molecule chosen from PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, CEACAM (e.g.,CEACAM-1, -3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/orTGFR beta. Inhibition of an inhibitory molecule can be performed byinhibition at the DNA, RNA or protein level. In embodiments, aninhibitory nucleic acid (e.g., a dsRNA, siRNA or shRNA), can be used toinhibit expression of an inhibitory molecule. In other embodiments, theinhibitor of an inhibitory signal is, a polypeptide e.g., a solubleligand, or an antibody or antigen-binding fragment thereof, that bindsto the inhibitory molecule. In one embodiment, the inhibitor is asoluble ligand (e.g., a CTLA-4-Ig), or an antibody or antibody fragmentthat binds to PD-L1, PD-L2 or CTLA-4. For example, the anti-TIM-3antibody molecule can be administered in combination with an anti-CTLA-4antibody, e.g., ipilimumab, for example, to treat a cancer (e.g., acancer chosen from: a melanoma, e.g., a metastatic melanoma; a lungcancer, e.g., a non-small cell lung carcinoma; or a prostate cancer). Inone embodiment, the anti-TIM-3 antibody molecule is administered aftertreatment with an anti-CTLA-4 antibody (e.g., ipilimumab) with orwithout a BRAF inhibitor (e.g., vemurafenib or dabrafenib).

In another embodiment, the anti-TIM-3 antibody molecule is administeredin combination with an anti-TIM-3 antibody or antigen-binding fragmentthereof.

In another embodiment, the anti-TIM-3 antibody molecule is administeredin combination with an anti-PD-1 antibody or antigen-binding fragmentthereof.

In yet other embodiments, the anti-TIM-3 antibody molecule isadministered in combination with an anti-TIM-3 antibody and ananti-TIM-3 antibody (or antigen-binding fragments thereof).

In another embodiment, the anti-TIM-3 antibody molecule is administeredin combination with a CEACAM inhibitor (e.g., CEACAM-1, -3 and/or -5inhibitor), e.g., an anti-CEACAM antibody molecule. In anotherembodiment, the anti-TIM-3 antibody molecule is administered incombination with a CEACAM-1 inhibitor, e.g., an anti-CEACAM-1 antibodymolecule. In another embodiment, the anti-TIM-3 antibody molecule isadministered in combination with a CEACAM-3 inhibitor, e.g., ananti-CEACAM-3 antibody molecule. In another embodiment, the anti-TIM-31antibody molecule is administered in combination with a CEACAM-5inhibitor, e.g., an anti-CEACAM-5 antibody molecule.

The combination of antibodies recited herein can be administeredseparately, e.g., as separate antibodies or antigen-binding fragmentsthereof, or linked, e.g., as a bispecific or trispecific antibodymolecule. In one embodiment, a bispecific antibody that includes ananti-TIM-3 antibody molecule and an anti-PD-1, anti-CEACAM (e.g.,anti-CEACAM-1, -3 and/or -5), or anti-TIM-3 antibody, or anantigen-binding fragment thereof, is administered. In certainembodiments, the combination of antibodies recited herein is used totreat a cancer, e.g., a cancer as described herein (e.g., a solid tumoror a hematologic malignancy).

In other embodiments, the anti-TIM-3 antibody molecule is administeredin combination with a cytokine. The cytokine can be administered as afusion molecule to the anti-TIM-3 antibody molecule, or as separatecompositions. In one embodiment, the anti-TIM-3 antibody is administeredin combination with one, two, three or more cytokines, e.g., as a fusionmolecule or as separate compositions. In one embodiment, the cytokine isan interleukin (IL) chosen from one, two, three or more of IL-1, IL-2,IL-12, IL-15 or IL-21. In one embodiment, a bispecific antibody moleculehas a first binding specificity to a first target (e.g., to TIM-3), asecond binding specificity to a second target (e.g., LAG-3 or PD-1), andis optionally linked to an interleukin (e.g., IL-12) domain e.g., fulllength IL-12 or a portion thereof. In certain embodiments, thecombination of anti-TIM-3 antibody molecule and the cytokine describedherein is used to treat a cancer, e.g., a cancer as described herein(e.g., a solid tumor).

In certain embodiments, the anti-TIM-3 antibody molecule is administeredin combination with an antibody specific against an HLA C, e.g., anantibody specific to Killer-cell Immunoglobulin-like Receptors (alsoreferred to herein as an “anti-KIR antibody”). In certain embodiments,the combination of anti-TIM-3 antibody molecule and anti-KIR antibody isused to treat a cancer, e.g., a cancer as described herein (e.g., asolid tumor, e.g., an advanced solid tumor).

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with a cellular immunotherapy (e.g., Provenge® (e.g.,Sipuleucel-T)), and optionally in combination with cyclophosphamide. Incertain embodiments, the combination of anti-TIM-3 antibody molecule,Provenge® and/or cyclophosphamide is used to treat a cancer, e.g., acancer as described herein (e.g., a prostate cancer, e.g., an advancedprostate cancer).

In another embodiment, the anti-TIM-3 antibody molecule is administeredin combination with a vaccine, e.g., a cancer vaccine, (e.g., adendritic cell renal carcinoma (DC-RCC) vaccine). In one embodiment, thevaccine is peptide-based, DNA-based, RNA-based, or antigen-based, or acombination thereof. In embodiments, the vaccine comprises one or morepeptides, nucleic acids (e.g., DNA or RNA), antigens, or a combinationthereof. In certain embodiments, the combination of anti-TIM-3 antibodymolecule and the DC-RCC vaccine is used to treat a cancer, e.g., acancer as described herein (e.g., a renal carcinoma, e.g., metastaticrenal cell carcinoma (RCC) or clear cell renal cell carcinoma (CCRCC)).

In another embodiment, the anti-TIM-3 antibody molecule is administeredin combination with an adjuvant.

In yet another embodiment, the anti-TIM-3 antibody molecule isadministered in combination with chemotherapy, and/or immunotherapy. Forexample, the anti-TIM-3 antibody molecule can be used to treat amyeloma, alone or in combination with one or more of: chemotherapy orother anti-cancer agents (e.g., thalidomide analogs, e.g.,lenalidomide), an anti-PD-1 antibody, tumor antigen-pulsed dendriticcells, fusions (e.g., electrofusions) of tumor cells and dendriticcells, or vaccination with immunoglobulin idiotype produced by malignantplasma cells. In one embodiment, the anti-TIM-3 antibody molecule isused in combination with an anti-TIM-3 antibody to treat a myeloma,e.g., a multiple myeloma.

In one embodiment, the anti-TIM-3 antibody molecule is used incombination with chemotherapy to treat a lung cancer, e.g., non-smallcell lung cancer. In one embodiment, the anti-TIM-3 antibody molecule isused with standard lung, e.g., NSCLC, chemotherapy, e.g., platinumdoublet therapy, to treat lung cancer. In yet other embodiments, theanti-TIM-3 antibody molecule is used in combination with anindoleamine-pyrrole 2,3-dioxygenase (IDO) inhibitor (e.g., INCB24360) ina subject with advanced or metastatic cancer (e.g., a patient withmetastic and recurrent NSCLC cancer).

In yet other embodiments, the anti-TIM-3 antibody molecule is used incombination with one or more of: an immune-based strategy (e.g.,interleukin-2 or interferon-α), a targeting agent (e.g., a VEGFinhibitor such as a monoclonal antibody to VEGF); a VEGF tyrosine kinaseinhibitor such as sunitinib, sorafenib, axitinib and pazopanib; an RNAiinhibitor; or an inhibitor of a downstream mediator of VEGF signaling,e.g., an inhibitor of the mammalian target of rapamycin (mTOR), e.g.,everolimus and temsirolimus. Any of such combinations can be used totreat a renal cancer, e.g., renal cell carcinoma (RCC) (e.g., clear cellrenal cell carcinoma (CCRCC)) or metastatic RCC.

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used in combinationwith a MEK inhibitor (e.g., a MEK inhibitor as described herein). Insome embodiments, the combination of the anti-TIM-3 antibody and the MEKinhibitor is used to treat a cancer (e.g., a cancer described herein).In some embodiments, the cancer treated with the combination is chosenfrom a melanoma, a colorectal cancer, a non-small cell lung cancer, anovarian cancer, a breast cancer, a prostate cancer, a pancreatic cancer,a hematological malignancy or a renal cell carcinoma. In certainembodiments, the cancer includes a BRAF mutation (e.g., a BRAF V600Emutation), a BRAF wildtype, a KRAS wildtype or an activating KRASmutation. The cancer may be at an early, intermediate or late stage.

In another embodiment, the anti-TIM-3 antibody molecule is used incombination with one, two or all of oxaliplatin, leucovorin or 5-FU(e.g., a FOLFOX co-treatment). Alternatively or in combination,combination further includes a VEGF inhibitor (e.g., a VEGF inhibitor asdisclosed herein). In some embodiments, the combination of theanti-TIM-3 antibody, the FOLFOX co-treatment, and the VEGF inhibitor isused to treat a cancer (e.g., a cancer described herein). In someembodiments, the cancer treated with the combination is chosen from amelanoma, a colorectal cancer, a non-small cell lung cancer, an ovariancancer, a breast cancer, a prostate cancer, a pancreatic cancer, ahematological malignancy or a renal cell carcinoma. The cancer may be atan early, intermediate or late stage.

In other embodiments, the anti-TIM-3 antibody molecule is administeredwith a tyrosine kinase inhibitor (e.g., axitinib) to treat renal cellcarcinoma and other solid tumors.

In other embodiments, the anti-TIM-3 antibody molecule is administeredwith a 4-1BB receptor targeting agent (e.g., an antibody that stimulatessignaling through 4-1BB (CD-137), e.g., PF-2566). In one embodiment, theanti-TIM-3 antibody molecule is administered in combination with atyrosine kinase inhibitor (e.g., axitinib) and a 4-1BB receptortargeting agent.

The anti-TIM-3 antibody molecule can be bound to a substance, e.g., acytotoxic agent or moiety (e.g., a therapeutic drug; a compound emittingradiation; molecules of plant, fungal, or bacterial origin; or abiological protein (e.g., a protein toxin) or particle (e.g., arecombinant viral particle, e.g., via a viral coat protein). Forexample, the antibody can be coupled to a radioactive isotope such as anα-, β-, or γ-emitter, or a β- and γ-emitter.

Additional Combination Therapies

The methods and compositions described herein (e.g., anti-TIM-3antibodies and methods of using them) can be used in combination withother agents or therapeutic modalities, e.g., a second therapeutic agentchosen from one or more of the agents listed in Table 6. In oneembodiment, the methods described herein include administering to thesubject an anti-TIM-3 antibody molecule as described herein (optionallyin combination with one or more inhibitors of PD-1, PD-L1, PD-L2, LAG-3,CEACAM (e.g., CEACAM-1 and/or CEACAM-5), or CTLA-4)), further includeadministration of a second therapeutic agent chosen from one or more ofthe agents listed in Table 6, in an amount effective to treat or preventa disorder, e.g., a disorder as described herein, e.g., a cancer. Whenadministered in combination, the anti-TIM-3 antibody molecule, theadditional agent (e.g., second or third agent), or all, can beadministered in an amount or dose that is higher, lower or the same thanthe amount or dosage of each agent used individually, e.g., as amonotherapy. In certain embodiments, the administered amount or dosageof the anti-TIM-3 antibody, the additional agent (e.g., second or thirdagent), or all, is lower (e.g., at least 20%, at least 30%, at least40%, or at least 50%) than the amount or dosage of each agent usedindividually, e.g., as a monotherapy. In other embodiments, the amountor dosage of the anti-TIM-3 antibody, the additional agent (e.g., secondor third agent), or all, that results in a desired effect (e.g.,treatment of cancer) is lower (e.g., at least 20%, at least 30%, atleast 40%, or at least 50% lower).

In other embodiments, the second therapeutic agent is chosen from one ormore of the agents listed in Table 6. In one embodiment, the cancer ischosen from a lung cancer (e.g., a non-small cell lung cancer (NSCLC)(e.g., a NSCLC with squamous and/or non-squamous histology, or a NSCLCadenocarcinoma), or disclosed in a publication listed in Table 6. Insome embodiments, the second therapeutic agent is chosen from one ormore of: 1) a protein kinase C (PKC) inhibitor; 2) a heat shock protein90 (HSP90) inhibitor; 3) an inhibitor of a phosphoinositide 3-kinase(PI3K) and/or target of rapamycin (mTOR); 4) an inhibitor of cytochromeP450 (e.g., a CYP17 inhibitor or a 17alpha-Hydroxylase/C17-20 Lyaseinhibitor); 5) an iron chelating agent; 6) an aromatase inhibitor; 7) aninhibitor of p53, e.g., an inhibitor of a p53/Mdm2 interaction; 8) anapoptosis inducer; 9) an angiogenesis inhibitor; 10) an aldosteronesynthase inhibitor; 11) a smoothened (SMO) receptor inhibitor; 12) aprolactin receptor (PRLR) inhibitor; 13) a Wnt signaling inhibitor; 14)a CDK4/6 inhibitor; 15) a fibroblast growth factor receptor 2(FGFR2)/fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) aninhibitor of macrophage colony-stimulating factor (M-CSF); 17) aninhibitor of one or more of c-KIT, histamine release, Flt3 (e.g.,FLK2/STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (e.g.,FLK-1/KDR), PDGFRbeta, c-KIT or Raf kinase C; 19) a somatostatin agonistand/or a growth hormone release inhibitor; 20) an anaplastic lymphomakinase (ALK) inhibitor; 21) an insulin-like growth factor 1 receptor(IGF-1R) inhibitor; 22) a P-Glycoprotein 1 inhibitor; 23) a vascularendothelial growth factor receptor (VEGFR) inhibitor; 24) a BCR-ABLkinase inhibitor; 25) an FGFR inhibitor; 26) an inhibitor of CYP11B2;27) a HDM2 inhibitor, e.g., an inhibitor of the HDM2-p53 interaction;28) an inhibitor of a tyrosine kinase; 29) an inhibitor of c-MET; 30) aninhibitor of JAK; 31) an inhibitor of DAC; 32) an inhibitor of11β-hydroxylase; 33) an inhibitor of IAP; 34) an inhibitor of PIMkinase; 35) an inhibitor of Porcupine; 36) an inhibitor of BRAF, e.g.,BRAF V600E or wild-type BRAF; 37) an inhibitor of HER3; 38) an inhibitorof MEK; or 39) an inhibitor of a lipid kinase, e.g., as described hereinand in Table 6.

In one embodiment, the second therapeutic agent is chosen from one ormore of: Compound A8, Compound A17, Compound A23, Compound A24, CompoundA27, Compound A29, Compound A33, and Compound A13.

In other embodiments, the second therapeutic agent is chosen from one ormore of: Compound A5, Compound A8, Compound A17, Compound A23, CompoundA24, Compound A29, and Compound A40.

In other embodiments, the second therapeutic agent is chosen from one ormore of: Compound A9, Compound A16, Compound A17, Compound A21, CompoundA22, Compound A25, Compound 28, Compound A48, and Compound 49.

In other embodiments, the second therapeutic agent is chosen from amodulator of an apoptotic pathway, e.g., an IDH1 inhibitor, or a Bcl-2or Bcl-XL inhibitor. In one embodiment, the second therapeutic agent ischosen from Compound A21, A14 or a combination thereof. Without beingbound by theory, TIM-3 is known to interact with PtdSer, which tends tobe exposed on the surface of apoptotic cells, and can causeimmunosuppression. Blockade of a PtdSer-TIM-3 interaction, e.g., usingan anti-TIM-3 antibody molecule as described herein may ameliorate orovercome the immunosuppression.

In other embodiments, the second therapeutic agent is an inhibitor ofCSF-1R, e.g., an anti-CSF-1R antibody or small molecule inhibitor (suchas Compound A15 or A33). These second therapeutic agents may inhibitmacrophages (e.g., M2 macrophages). In certain embodiments, such secondtherapeutic agents can facilitate the conversion to M1 macrophages.

In embodiments, the second therapeutic agent is administered at atherapeutic or lower-than therapeutic dose. In certain embodiments, theconcentration of the second therapeutic agent that is required toachieve inhibition, e.g., growth inhibition, is lower when the secondtherapeutic agent is administered in combination with the anti-TIM-3antibody molecule than when the second therapeutic agent is administeredindividually. In certain embodiments, the concentration of theanti-TIM-3 antibody molecule that is required to achieve inhibition,e.g., growth inhibition, is lower when the anti-TIM-3 antibody moleculeis administered in combination with the second therapeutic agent thanwhen the anti-TIM-3 antibody molecule is administered individually. Incertain embodiments, in a combination therapy, the concentration of thesecond therapeutic agent that is required to achieve inhibition, e.g.,growth inhibition, is lower than the therapeutic dose of the secondtherapeutic agent as a monotherapy, e.g., 10-20%, 20-30%, 30-40%,40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower. In certain embodiments,in a combination therapy, the concentration of the anti-TIM-3 antibodymolecule that is required to achieve inhibition, e.g., growthinhibition, is lower than the therapeutic dose of the anti-TIM-3antibody molecule as a monotherapy, e.g., 10-20%, 20-30%, 30-40%,40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower.

Detection

In some aspects, the present disclosure provides methods for detectingthe presence of TIM-3 in a sample, e.g., in vitro or in vivo (e.g., abiological sample, e.g., blood, serum, semen or urine, or a tissuebiopsy, e.g., from a hyperproliferative or cancerous lesion). Themethods herein can be used to evaluate (e.g., monitor treatment orprogression of, diagnose and/or stage a disorder described herein, e.g.,an immune disorder, a cancer, or an infectious disease, in a subject).The method may include: (i) contacting the sample with (and optionally,a reference, e.g., a control sample), or administering to the subject,an anti-TIM-3 antibody molecule as described herein, under conditionsthat allow interaction to occur, and (ii) detecting whether there isformation of a complex between the antibody molecule and the sample (andoptionally, the reference, e.g., control, sample). Formation of thecomplex is indicative of the presence of TIM-3, and can indicate thesuitability or need for a treatment described herein. The method caninvolve, e.g., an immunohistochemistry, immunocytochemistry, flowcytometry, antibody molecule complexed magnetic beads, ELISA assays,PCR-techniques (e.g., RT-PCR).

Typically, the anti-TIM-3 antibody molecule used in the in vivo and invitro diagnostic methods is directly or indirectly labeled with adetectable substance to facilitate detection of the bound or unboundbinding agent. Suitable detectable substances include variousbiologically active enzymes, prosthetic groups, fluorescent materials,luminescent materials, paramagnetic (e.g., nuclear magnetic resonanceactive) materials, and radioactive materials.

Additional embodiments provide a method of treating a cancer,comprising: identifying in a sample (e.g., a subject's sample comprisingcancer cells and optionally immune cells such as TILs) the presence ofone, two or all of PD-L1, CD8, or IFN-γ, thereby providing a value forone, two or all of PD-L1, CD8, and IFN-γ. The method can further includecomparing the PD-L1, CD8, and/or IFN-γ values to a reference value,e.g., a control value. If the PD-L1, CD8, and/or IFN-γ values aregreater than the reference value, e.g., the control values,administering a therapeutically effective amount of an anti-TIM-3antibody (e.g., an anti-TIM-3 antibody described herein) to the subject,optionally in combination with one or more other agents, therebytreating the cancer. The cancer may be, e.g., a cancer described herein,such as lung cancer (squamous), lung cancer (adenocarcinoma), head andneck cancer, cervical cancer (squamous), stomach cancer, thyroid cancer,melanoma, nasopharyngeal cancer, or breast cancer, e.g., TN breastcancer, e.g., IM-TN breast cancer. In some embodiments, the cancer isER+ breast cancer or pancreatic cancer.

Also provided is a method of treating a cancer, comprising: testing asample (e.g., a subject's sample comprising cancer cells) for thepresence of PD-L1, thereby identifying a PD-L1 value, comparing thePD-L1 value to a control value, and if the PD-L1 value is greater thanthe control value, administering a therapeutically effective amount ofan anti-TIM-3 antibody (e.g., an anti-TIM-3 antibody described herein)to the subject, optionally in combination with one or more other agents,e.g., an anti-PD-1 antibody molecule, thereby treating the cancer. Thecancer may be, e.g., a cancer as described herein, such as cancer isnon-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLC squamous cellcarcinoma (SCC), or hepatocellular carcinoma (HCC).

In some aspects, the present disclosure provides diagnostic ortherapeutic kits that include the anti-TIM-3 antibody moleculesdescribed herein and instructions for use.

The disclosure contemplates all combinations of any one or more of theforegoing aspects and/or embodiments, as well as combinations with anyone or more of the embodiments set forth in the detailed description andexamples.

Other features, objects, and advantages of the compositions and methodsherein will be apparent from the description and drawings, and from theclaims.

Figures and Tables are provided herewith.

BRIEF DESCRIPTION OF DRAWINGS

Each of the Figures is described herein in more detail.

FIGS. 1A-1B depict exemplary anti-TIM-3 antibodies. FIG. 1A provides theheavy chain and light chain variable regions of ABTIM3 (SEQ ID NOS: 1and 2, respectively, in order of appearance). FIG. 1B provides asequence alignment between the variable regions of ABTIM3 and murine(mouse) germline antibodies (SEQ ID NOS: 134 and 135, respectively, inorder of appearance). The CDRs are boxed (depicted in white text on ablack background in the priority documents).

FIGS. 2A-2E illustrate the binding and activity of various anti-TIM-3antibodies. FIG. 2A summarizes affinity data for the murine antibodyABTIM3 and another TIM-3 binding antibody. FIG. 2B shows a binding curveof one panel of antibodies for human TIM-3 in transfected cells. FIG. 2Cshows a binding curve of a second panel of antibodies, including ABTIM3(triangles) for human TIM-3 in transfected cells. FIG. 2D shows abinding curve of ABTIM3 and other anti-TIM-3 antibodies for cynomolgusmonkey TIM-3. FIG. 2E shows the affinity of several anti-TIM-3antibodies for cynomolgus monkey TIM-3. Monoclonal antibody ABTIM3 hasthe highest affinity of the antibodies tested in these experiments,indicating it has good cross-reactivity with human and monkey targets.

FIGS. 3A-3B show that anti-TIM-3 monoclonal antibodies, includingandABTIM3, bind to the IgV domain, while 4A4 binds to the mucin domain.FIG. 3A illustrates the recombinant construct used for epitope analysis.FIG. 3B shows that the anti-TIM-3 monoclonal antibody (anti-TIM-3 #3),and anti-PD-L1 control monoclonal antibodies (anti-PD-L1 #1 and #2),bind to the chimeric protein of FIG. 3A, while anti-TIM-3 #2 and ABTIM3do not substantially bind.

FIG. 4 illustrates that anti-TIM-3 antibodies anti-TIM-3 #2 and ABTIM3block binding of TIM-3 to PtdSer (phosphatidylserine).

FIGS. 5A-5B illustrate that the anti-TIM-3 antibody ABTIM3 enhancesIFN-gamma secretion and proliferation in IL-12 Stimulated CD4+ T Cells.FIG. 5A shows the results of a representative experiment where cellswere exposed to antibodies ABTIM3, anti-TIM-3 #2, mIgG1, and anti-PD-L1control antibody (from left to right). IFN-gamma levels were measured byflow cytometry. FIG. 5B quantifies IFN-gamma expression in cells exposedto these four antibodies.

FIG. 6 shows that a ABTIM3 blockade enhances in vitro cytotoxic activityof purified NK cells.

FIG. 7 shows that humanized anti-TIM-3 antibodies competed for bindingwith the parent murine ABTIM3 antibody in a FACS assay.

FIGS. 8A-8B illustrate that humanized anti-TIM-3 antibodies bind tocells expressing human TIM-3. FIG. 8A shows that humanized anti-TIM-3antibodies bound to cells expressing huTIM-3 in a FACS assay. FIG. 8Bshows that the humanized anti-TIM-3 antibodies competed with theparental murine ABTIM3 for cells expressing huTIM-3 in a FACS assay.

FIGS. 9A-9B illustrate the structure of ABTIM3-hum21 Fab binding toTIM-3. FIG. 9A shows the overall structure of ABTIM3-hum21 Fab bindingto TIM-3. Labeled in the figure are 1) the deduced PtdSer, Ca²⁺ andGalectin-9 binding sites on human TIM-3 and 2) names of the β strandsand BC, FG and CC′ loops. FIG. 9B shows a detailed view of ABTIM3-hum21epitope residues on TIM-3 (shown as sticks and labeled). FIG. 9Bdiscloses residues 56-61 (“GACPVF”) as SEQ ID NO: 136 and residues119-127 (“NDEKFNLKL”) as SEQ ID NO: 137.

FIGS. 10A-10C shows the comparison of ABTIM3-hum21 epitope withCEACAM-1-binding site on human TIM-3. FIG. 10A shows the comparison ofthe crucial CEACAM-1-binding residues of TIM-3 (residues 117-120(“IMND”) disclosed as SEQ ID NO: 138) (left panel, grey surface,residues are labeled) and the ABTIM3-hum21 epitope (right panel, greysurface, residues that overlap with CECAM1-binding site are labeled).Since TIM-3 is oriented the same way in both panels, it is obvious thatABTIM3-hum21 epitope overlaps with CEACAM-1 binding site. FIG. 10B showsthe K122 of TIM-3 forms hydrogen bond with CEACAM-1 (left panel), and iscompleted blocked by ABTIM3-hum21 (right panel). FIG. 10C showstwo-angle views of the superimposition of TIM-3/ABTIM3-hum21 Fab andTIM-3/CEACAM-1 structures, which shows significant clash betweenABTIM3-hum21 and TIM-3, indicating ABTIM3-hum21 will disrupt CEACAM-1binding to TIM-3.

FIG. 11 illustrates the comparison of PtdSer-mediated membranepenetration of moue TIM-3 (left panel) and binding of ABTIM3-hum21 tohuman TIM-3 (right panel). The two TIM-3 structures are oriented thesame way. The attacking angle of ABTIM3-hum21 is similar to theorientation of the membrane penetrated by TIM-3, which suggests thatABTIM3-hum21 will prevent PtdSer-mediated penetration of TIM-3.

FIG. 12 shows the cancer indications with the highest expression ofTIM-3 (HAVCR2) from the TCGA database.

FIG. 13 shows the cancer indications with the highest expression of amacrophage expression signature from the TCGA database.

FIG. 14 shows exemplary cancers having relatively high proportions ofpatients that are triple-positive for PD-L1/CD8/IFN-γ.

FIG. 15 shows exemplary ER+ breast cancer and pancreatic cancer havingrelatively low proportions for patients that are triple positive forPD-L1/CD8/IFN-γ.

FIG. 16 shows the proportion of exemplary breast cancer patients thatare triple positive for PD-L1/CD8/IFN-γ.

FIG. 17 shows the proportion of exemplary colon cancer patients that aretriple positive for PD-L1/CD8/IFN-γ.

FIG. 18 shows the peptides that are monitored in HDx-MS experiments onthe human TIM-3 (residues 23 to 135(“SEVEYRAEVGQNAYLPCFYTPAAPGNLVPVCWGKGACPVFECGNVVLRTDERDVNYWTSRYWLNGDFRKGDVSLTIENVTLADSGIYCCRIQIPGIMNDEKFNLKLVIKPAKVT”) as SEQ IDNO: 139). Each bar represents a peptide.

FIG. 19 illustrates the difference in deuterium uptake for the TIM-3ABTIM3-hum03 complex (grey bars) and the TIM-3 ABTIM3-hum11 complex(black bars) for amino acids 22 through 127. All differences arerelative to the deuterium uptake of unbound TIM-3 (control).

FIG. 20 shows the competition between ABTIM3-hum21 and ABTIM3-hum03 andABTIM3-hum11 for binding to human TIM3, as determined by flow cytometryassay.

FIG. 21 shows a representative sensogram from a Biacore competitionassay testing the competition between a 1^(st) antibody and a 2^(nd)antibody for immobilized human TIM-3.

FIG. 22 shows that ABTIM3 increases proliferation in a co-culturecontaining dendritic cells and T cells (DC-T co-culture). DC-Tco-cultures were incubated with no antibody or a titrated dilutionseries (0.01-25 μg/mL) of the following antibodies mouse IgG1 (control),ABTIM3 or anti-TIM3 #3 antibody.

FIGS. 23A-23B show the concentration of ABTIM3-hum11 detected in theserum over time in rodents. The indicated dosages were injected intomice or rats, and the concentration of antibody in the blood wascalculated at the indicated time points. FIG. 23A shows the mean serumconcentration of BTIM3-hum11 in mice after antibody administration. FIG.23B shows the mean serum concentration of ABTIM3-hum11 in rats afterantibody administration.

BRIEF DESCRIPTION OF THE TABLES

Each of the Tables is described herein in more detail.

Table 1 summarizes the sequences of the murine anti-TIM-3 antibody,ABTIM3.

Table 2 depicts the amino acid sequences of ABTIM3 heavy chain variabledomain and light chain variable domain.

Table 3 depicts the amino acid sequences of ABTIM3 heavy chain CDRs andlight chain CDRs.

Table 4 is a summary of the amino acid and nucleotide sequences for themurine and humanized anti-TIM-3 antibody molecules. The antibodymolecules include murine ABTIM3 and humanized anti-TIM-3 antibodies:ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05,ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10,ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15,ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20,ABTIM3-hum21, ABTIM3-hum22, and ABTIM3-hum23. The amino acid andnucleotide sequences of the heavy and light chain CDRs, the amino acidand nucleotide sequences of the heavy and light chain variable regions,and the amino acid and nucleotide sequences of the heavy and lightchains are shown in this Table.

Table 5 depicts the constant region amino acid sequences of human IgGheavy chains and human kappa light chain.

Table 6 is a summary of selected therapeutic agents that can beadministered in combination with the anti-TIM-3 antibody molecules andother immunomodulators (e.g., one or more of: an activator of acostimulatory molecule and/or an inhibitor of an immune checkpointmolecule) described herein. Table 6 provides from left to right thefollowing: the Compound Designation of the second therapeutic agent, theCompound structure, and Patent publication(s) disclosing the Compound.

Table 7 summarizes the K_(D) values for anti-TIM-3 antibody binding toactivated PBMCs.

Table 8 summarizes the K_(D) values for anti-TIM-3 antibody binding toPD-L1 IgV/TIM-3 mucin construct.

Table 9 summarizes the K_(D) values for a panel of humanized anti-TIM-3antibodies as measured by Biacore assay.

Table 10 summarizes the K_(D) values for anti-TIM-3 antibody binding tocells expressing human TIM-3.

Table 11 summarizes the K_(D) values for anti-TIM-3 antibody binding toTIM-3-Ig.

Table 12 summarizes the amino acid sequences used for crystal structuredetermination.

Table 13 summarizes the amino acids in TIM-3 and anti-TIM-3 antibodythat participate in the binding interaction.

Table 14 summerizes the Biacore competition assay cycles.

Table 15 summerizes the results from Biacore competition assay.

Table 16 summerizes the pharmacokinetic properties of ABTIM3-hum11.

DETAILED DESCRIPTION

T-cell immunoglobulin domain and mucin domain 3 (TIM-3, also known asHepatitis A virus cellular receptor 2, and HAVCR2) is a cell surfaceprotein expressed, e.g., on activated CD4+ and CD8+ T cells, naturalregulatory T cells (nTregs), NK cells, and innate cells, e.g.,macrophages, monocytes and dendritic cells (DCs). TIM-3 is generally notexpressed on naïve T cells, but rather upregulated on activated,effector T cells, e.g., on a PD-1+ subset of cells. TIM-3 is alsoexpressed on tissue site natural regulatory cells and in murine models.TIM-3+ Tregs have been shown to have a more suppressive phenotype whileTIM-3+ Tregs have also been shown to correlate with disease severity inNSCLC, hepatocellular and ovarian carcinoma. TIM-3 is constitutivelyexpressed on DCs, monocytes/macrophages and NK cells, and blockade ofTIM-3 has been shown to correlate with increased cytotoxicity in NKcells; increased secretion of IL-12/TNF-α by monocytes/macrophages; andincreased NF-κB expression in DCs. Blockade of TIM-3 (partially aloneand additively or synergistically in combination with PD-1 pathwayblockade) has shown anti-tumor efficacy in several preclinical cancermodels, including CT26 colon carcinoma (Sakuishi et al., J Exp Med.2010; 207(10):2187-94), WT3 sarcoma and TRAMP-C1 prostate carcinoma(Ngiow et al., Cancer Res. 2011; 71(10):3540-3551). Recent studies havehighlighted TIM-3 as an important player in the T effector cellexhaustion and suppression that takes place in chronic immune conditionssuch as infection, e.g., bacterial or viral, and cancer in both humansand experimental models. TIM-3 has been described as an inhibitoryreceptor in the immunological synapse, and blocking of TIM-3 may enhanceimmune response against infection and cancer.

Blockade of TIM-3 has been shown to restore activity in effector cells,such as cytokine secretion and proliferation. In virally exhausted cellpopulations, e.g., cells infected with HCV, TIM-3-expressing cells(TIM3+ cells) express less TNF-alpha and IFN-gamma cytokines than TIM-3negative cells in both effector cell populations, CD4+ and CD8+ T cells(Golden-Mason et al., 2009, J. Virol, 83:9122). Blockade of TIM-3restores proliferation in CD8+ T cells from an HIV patient, or in cellsthat recapitulate viral exhaustion (Jones et al., 2008, J. Exp. Med.,205:2763), or proliferation and IFN-γ and/or TNF-α secretion in NY-ESO-1specific T cells from PBMCs from metastatic patients (Fourcade et al.,2010, J. Exp. Med., 207:2175). TIM-3 blockade may also diminish thesuppressor activity of regulatory T cells. TIM-3+ T cells have beenfound to be concentrated in tumors, and contribute to theimmunosuppressive tumor environment (Sakuishi et al., 2013,Oncoimmunology, 2:e23849; Gao et al., 2012, Plos One; and Yan et al.,2013, Plos One.). Thus, blockade of TIM-3, e.g., by antibodies thatinhibit TIM-3 function, can improve the immune response againstinfection and anti-tumor immunity.

TIM-3 has also been implicated in regulating immune response throughmacrophage activity. Blockade of TIM-3 leads to an increase inTLR-mediated IL-12 production (Zhang et al., 2010, J Leukoc Biol,91:189). Thus, TIM-3 blockade may increase immune stimulation propertiesof macrophages to enhance immune response against infection andanti-tumor activity.

TIM-3 has five reported ligands: Galectin-9 (Gal-9), phosphatidylserine(PtdSer), HMGB1, Semaphorin-4A, and CEACAM-1. S-type lectin galectin-9can inhibit TIM-3-associated Th1 effector function and induce apoptosison TIM-3-expressing T cells in murine models. PtdSer usually resides onthe intracellular side of the plasma membrane, but is flipped to theextracellular side during apoptosis. PtdSer binds a preserved cleft inall three human TIM family members (TIM-1, 3, 4). Inhibition of PtdSerbinding to TIM-3 may activate T-cell response. Galectin-9 is secreted bytumor cells and can contribute to evasion from anti-tumor immunity. DNAalarmin HMGB1, for which TIM-3 may act as a “sink,” can prevent theHMGB1/RAGE interactions that stimulate innate immunity. Semaphorin-4Aand CEACAM-1 (another immune checkpoint molecule whose inhibition canenhance immune response) can interact with TIM-3 both in cis as aheterodimer on T cells and in trans as a ligand. Interaction betweenCEACAM-1 and TIM-3 may help mediate block immune response signaling.Co-blockade of TIM-3 and CEACAM-1 in CT26 colon carcinoma showed similarefficacy to that seen for co-blockade of PD-L1 and TIM-3.

The TIM-3 cytoplasmic tail has seven sites for tyrosine phosphorylationand no known inhibitory (i.e., ITIM) motifs, which suggests that TIM-3could co-stimulate with the T cell receptor, leading to functionalexhaustion through increased T cell signaling. TIM-3 can interact withFyn and facilitate accumulation of receptor phosphatases CD148 and CD45at the immunologic synapse. The presence of CEACAM-1 as a co-receptor inthe TIM-3/CEACAM-1 heterodimer suggests that this co-expression may leadto inhibitory signaling in T cells via the ITIM motif in the CEACAM-1cytoplasmic tail which has been shown to interact with both SHP1 andSHP2.

Disclosed herein are antibody molecules that bind to TIM-3 with highaffinity and specificity. In one embodiment, humanized antibodiesagainst TIM-3 are disclosed. Additional aspects of the invention includenucleic acid molecules encoding the antibody molecules, expressionvectors, host cells and methods for making the antibody molecules arealso provided. Immunoconjugates, multi- or bispecific antibody moleculesand pharmaceutical compositions comprising the antibody molecules arealso provided. The anti-TIM-3 antibody molecules disclosed herein can beused (alone or in combination with other agents or therapeuticmodalities) to treat, prevent and/or diagnose immune disorders, cancer,infectious disease, Crohn's disease, sepsis, SIRS (Systemic InflammatoryResponse Syndrome), and glomerulonephritis. Thus, compositions andmethods for detecting TIM-3, as well as methods for treating variousdisorders, including cancer and immune disorders using the anti-TIM-3antibody molecules are disclosed herein.

The term “TIM-3” include isoforms, mammalian, e.g., human TIM-3, specieshomologs of human TIM-3, and analogs comprising at least one commonepitope with TIM-3. The amino acid sequence of TIM-3, e.g, human TIM-3,is known in the art, e.g., Sabatos et al., 2003. Nat Immunol,4(11):1102.

Definitions

As used herein, the articles “a” and “an” refer to one or to more thanone (e.g., to at least one) of the grammatical object of the article.

The term “or” is used herein to mean, and is used interchangeably with,the term “and/or”, unless context clearly indicates otherwise.

“About” and “approximately” shall generally mean an acceptable degree oferror for the quantity measured given the nature or precision of themeasurements. Exemplary degrees of error are within 20 percent (%),typically, within 10%, and more typically, within 5% of a given value orrange of values.

The compositions and methods disclosed herein encompass polypeptides andnucleic acids having the sequences specified, or sequences substantiallyidentical or similar thereto, e.g., sequences at least 85%, 90%, 95%identical or higher to the sequence specified. In the context of anamino acid sequence, the term “substantially identical” is used hereinto refer to a first amino acid that contains a sufficient or minimumnumber of amino acid residues that are i) identical to, or ii)conservative substitutions of aligned amino acid residues in a secondamino acid sequence such that the first and second amino acid sequencescan have a common structural domain and/or common functional activity.For example, amino acid sequences that contain a common structuraldomain having at least about 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%,97%, 98% or 99% identity to a reference sequence, e.g., a sequenceprovided herein.

In the context of nucleotide sequence, the term “substantiallyidentical” is used herein to refer to a first nucleic acid sequence thatcontains a sufficient or minimum number of nucleotides that areidentical to aligned nucleotides in a second nucleic acid sequence suchthat the first and second nucleotide sequences encode a polypeptidehaving common functional activity, or encode a common structuralpolypeptide domain or a common functional polypeptide activity. Forexample, nucleotide sequences having at least about 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence,e.g., a sequence provided herein.

The term “functional variant” refers polypeptides that have asubstantially identical amino acid sequence to the naturally-occurringsequence, or are encoded by a substantially identical nucleotidesequence, and are capable of having one or more activities of thenaturally-occurring sequence.

To determine the percent identity of two amino acid sequences, or of twonucleic acid sequences, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in one or both of a first and asecond amino acid or nucleic acid sequence for optimal alignment andnon-homologous sequences can be disregarded for comparison purposes). Ina preferred embodiment, the length of a reference sequence aligned forcomparison purposes is at least 30%, e.g., at least 40%, 50%, 60%, e.g.,at least 70%, 80%, 90%, 100% of the length of the reference sequence.The amino acid residues or nucleotides at corresponding amino acidpositions or nucleotide positions are then compared. When a position inthe first sequence is occupied by the same amino acid residue ornucleotide as the corresponding position in the second sequence, thenthe molecules are identical at that position.

The percent identity between the two sequences is a function of thenumber of identical positions shared by the sequences, taking intoaccount the number of gaps, and the length of each gap, which need to beintroduced for optimal alignment of the two sequences.

The comparison of sequences and determination of percent identitybetween two sequences can be accomplished using a mathematicalalgorithm. In a some embodiments, the percent identity between two aminoacid sequences is determined using the Needleman and Wunsch ((1970) J.Mol. Biol. 48:444-453) algorithm which has been incorporated into theGAP program in the GCG software package (available athttp://www.gcg.com), using either a Blossum 62 matrix or a PAM250matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a lengthweight of 1, 2, 3, 4, 5, or 6. In certain embodiments, the percentidentity between two nucleotide sequences is determined using the GAPprogram in the GCG software package (available at http://www.gcg.com),using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80and a length weight of 1, 2, 3, 4, 5, or 6. One suitable set ofparameters (and the one that should be used unless otherwise specified)are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extendpenalty of 4, and a frameshift gap penalty of 5.

The percent identity between two amino acid or nucleotide sequences canbe determined using the algorithm of E. Meyers and W. Miller ((1989)CABIOS, 4:11-17) which has been incorporated into the ALIGN program(version 2.0), using a PAM120 weight residue table, a gap length penaltyof 12 and a gap penalty of 4.

The nucleic acid and protein sequences described herein can be used as a“query sequence” to perform a search against public databases to, forexample, identify other family members or related sequences. Suchsearches can be performed using the NBLAST and XBLAST programs (version2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLASTnucleotide searches can be performed with the NBLAST program, score=100,wordlength=12 to obtain nucleotide sequences homologous to a nucleicacid as described herein. BLAST protein searches can be performed withthe XBLAST program, score=50, wordlength=3 to obtain amino acidsequences homologous to protein molecules described herein. To obtaingapped alignments for comparison purposes, Gapped BLAST can be utilizedas described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402.When utilizing BLAST and Gapped BLAST programs, the default parametersof the respective programs (e.g., XBLAST and NBLAST) can be used. Seewww.ncbi.nlm.nih.gov.

As used herein, the term “hybridizes under low stringency, mediumstringency, high stringency, or very high stringency conditions”describes conditions for hybridization and washing. Guidance forperforming hybridization reactions can be found in Current Protocols inMolecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which isincorporated by reference. Aqueous and nonaqueous methods are describedin that reference and either can be used. Specific hybridizationconditions referred to herein are as follows: 1) low stringencyhybridization conditions in 6× sodium chloride/sodium citrate (SSC) atabout 45° C., followed by two washes in 0.2×SSC, 0.1% SDS at least at50° C. (the temperature of the washes can be increased to 55° C. for lowstringency conditions); 2) medium stringency hybridization conditions in6×SSC at about 45° C., followed by one or more washes in 0.2×SSC, 0.1%SDS at 60° C.; 3) high stringency hybridization conditions in 6×SSC atabout 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65°C.; and preferably 4) very high stringency hybridization conditions are0.5M sodium phosphate, 7% SDS at 65° C., followed by one or more washesat 0.2×SSC, 1% SDS at 65° C. Very high stringency conditions (4) aresuitable conditions and the ones that should be used unless otherwisespecified.

It is understood that the molecules described herein may have additionalconservative or non-essential amino acid substitutions, which do nothave a substantial effect on their functions.

A “conservative amino acid substitution” is one in which the amino acidresidue is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art. These families include amino acids with basicside chains (e.g., lysine, arginine, histidine), acidic side chains(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine),nonpolar side chains (e.g., alanine, valine, leucine, isoleucine,proline, phenylalanine, methionine, tryptophan), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine).

The terms “polypeptide”, “peptide” and “protein” (if single chain) areused interchangeably herein.

The terms “nucleic acid,” “nucleic acid sequence,” “nucleotidesequence,” or “polynucleotide sequence,” and “polynucleotide” are usedinterchangeably.

The term “isolated,” as used herein, refers to material that is removedfrom its original or native environment (e.g., the natural environmentif it is naturally occurring). For example, a naturally-occurringpolynucleotide or polypeptide present in a living animal is notisolated, but the same polynucleotide or polypeptide, separated by humanintervention from some or all of the co-existing materials in thenatural system, is isolated. Such polynucleotides could be part of avector and/or such polynucleotides or polypeptides could be part of acomposition, and still be isolated in that such vector or composition isnot part of the environment in which it is found in nature.

Various aspects of the compositions and methods herein are described infurther detail below. Additional definitions are set out throughout thespecification.

Antibody Molecules

In some embodiments, the antibody molecule binds to a mammalian, e.g.,human, TIM-3. For example, the antibody molecule binds specifically toan epitope, e.g., linear or conformational epitope, (e.g., an epitope asdescribed herein) on TIM-3. In some embodiments, the epitope is at leasta portion of the IgV domain of human or cynomolgus TIM-3.

As used herein, the term “antibody molecule” refers to a protein, e.g.,an immunoglobulin chain or fragment thereof, comprising at least oneimmunoglobulin variable domain sequence. The term “antibody molecule”includes, for example, a monoclonal antibody (including a full lengthantibody which has an immunoglobulin Fc region). In an embodiment, anantibody molecule comprises a full length antibody, or a full lengthimmunoglobulin chain. In an embodiment, an antibody molecule comprisesan antigen bindng or functional fragment of a full length antibody, or afull length immunoglobulin chain.

In an embodiment, an antibody molecule is a monospecific antibodymolecule and binds a single epitope. E.g., a monospecific antibodymolecule having a plurality of immunoglobulin variable domain sequences,each of which binds the same or substantially the same epitope.

In an embodiment, an antibody molecule is a multispecific antibodymolecule, e.g., it comprises a plurality of immunoglobulin variabledomains sequences, wherein a first immunoglobulin variable domainsequence of the plurality has binding specificity for a first epitopeand a second immunoglobulin variable domain sequence of the pluralityhas binding specificity for a second epitope. In an embodiment, thefirst and second epitopes are on the same antigen, e.g., the sameprotein (or subunit of a multimeric protein). In an embodiment the firstand second epitopes overlap or substantially overlap. In an embodiment,the first and second epitopes do not overlap or do not substantiallyoverlop. In an embodiment, the first and second epitopes are ondifferent antigens, e.g., the different proteins (or different subunitsof a multimeric protein). In an embodiment, a multispecific antibodymolecule comprises a third, fourth or fifth immunoglobulin variabledomain. In an embodiment, a multispecific antibody molecule is abispecific antibody molecule, a trispecific antibody molecule, ortetraspecific antibody molecule,

In an embodiment, a multispecific antibody molecule is a bispecificantibody molecule. A bispecific antibody has specificity for no morethan two antigens. A bispecific antibody molecule is characterized by afirst immunoglobulin variable domain sequence which has bindingspecificity for a first epitope and a second immunoglobulin variabledomain sequence that has binding specificity for a second epitope. In anembodiment, the first and second epitopes are on the same antigen, e.g.,the same protein (or subunit of a multimeric protein). In an embodiment,the first and second epitopes overlap or substantially overlap. In anembodiment the first and second epitopes do not overlap or do notsubstantially overlap. In an embodiment the first and second epitopesare on different antigens, e.g., the different proteins (or differentsubunits of a multimeric protein). In an embodiment a bispecificantibody molecule comprises a heavy chain variable domain sequence and alight chain variable domain sequence which have binding specificity fora first epitope and a heavy chain variable domain sequence and a lightchain variable domain sequence which have binding specificity for asecond epitope. In an embodiment, a bispecific antibody moleculecomprises a half antibody having binding specificity for a first epitopeand a half antibody having binding specificity for a second epitope. Inan embodiment, a bispecific antibody molecule comprises a half antibody,or fragment thereof, having binding specificity for a first epitope anda half antibody, or fragment thereof, having binding specificity for asecond epitope. In an embodiment a bispecific antibody moleculecomprises a scFv, or fragment thereof, have binding specificity for afirst epitope and a scFv, or fragment thereof, have binding specificityfor a second epitope. In an embodiment the first epitope is located onTIM-3 and the second epitope is located on a PD-1, LAG-3, CEACAM (e.g.,CEACAM-1, CEACAM-3 and/or CEACAM-5), PD-L1, or PD-L2.

In an embodiment, an antibody molecule comprises a diabody, and asingle-chain molecule, as well as an antigen-binding fragment of anantibody (e.g., Fab, F(ab′)₂, and Fv). For example, an antibody moleculecan include a heavy (H) chain variable domain sequence (abbreviatedherein as VH), and a light (L) chain variable domain sequence(abbreviated herein as VL). In an embodiment an antibody moleculecomprises or consists of a heavy chain and a light chain (referred toherein as a half antibody. In another example, an antibody moleculeincludes two heavy (H) chain variable domain sequences and two light (L)chain variable domain sequence, thereby forming two antigen bindingsites, such as Fab, Fab′, F(ab′)₂, Fc, Fd, Fd′, Fv, single chainantibodies (scFv for example), single variable domain antibodies,diabodies (Dab) (bivalent and bispecific), and chimeric (e.g.,humanized) antibodies, which may be produced by the modification ofwhole antibodies or those synthesized de novo using recombinant DNAtechnologies. These functional antibody fragments retain the ability toselectively bind with their respective antigen or receptor. Antibodiesand antibody fragments can be from any class of antibodies including,but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass(e.g., IgG1, IgG2, IgG3, and IgG4) of antibodies. The preparation ofantibody molecules can be monoclonal or polyclonal. An antibodymoleculecan also be a human, humanized, CDR-grafted, or in vitro generatedantibody. The antibody can have a heavy chain constant region chosenfrom, e.g., IgG1, IgG2, IgG3, or IgG4. The antibody can also have alight chain chosen from, e.g., kappa or lambda. The term“immunoglobulin” (Ig) is used interchangeably with the term “antibody”herein.

Examples of antigen-binding fragments of an antibody molecule include:(i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CLand CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprisingtwo Fab fragments linked by a disulfide bridge at the hinge region;(iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fvfragment consisting of the VL and VH domains of a single arm of anantibody, (v) a diabody (dAb) fragment, which consists of a VH domain;(vi) a camelid or camelized variable domain; (vii) a single chain Fv(scFv), see e.g., Bird et al. (1988) Science 242:423-426; and Huston etal. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) a singledomain antibody. These antibody fragments may be obtained using anysuitable method, including several conventional techniques known tothose with skill in the art, and the fragments can be screened forutility in the same manner as are intact antibodies.

The term “antibody” includes intact molecules as well as functionalfragments thereof. Constant regions of the antibodies can be altered,e.g., mutated, to modify the properties of the antibody (e.g., toincrease or decrease one or more of: Fc receptor binding, antibodyglycosylation, the number of cysteine residues, effector cell function,or complement function).

The antibodies disclosed herein can also be single domain antibodies.Single domain antibodies can include antibodies whose complementarydetermining regions are part of a single domain polypeptide. Examplesinclude, but are not limited to, heavy chain antibodies, antibodiesnaturally devoid of light chains, single domain antibodies derived fromconventional 4-chain antibodies, engineered antibodies and single domainscaffolds other than those derived from antibodies. Single domainantibodies may be any of the art, or any future single domainantibodies. Single domain antibodies may be derived from any speciesincluding, but not limited to mouse, human, camel, llama, fish, shark,goat, rabbit, and bovine. According to some aspects, a single domainantibody is a naturally occurring single domain antibody known as heavychain antibody devoid of light chains. Such single domain antibodies aredisclosed in WO 9404678, for example. For clarity reasons, this variabledomain derived from a heavy chain antibody naturally devoid of lightchain is known herein as a VHH or nanobody to distinguish it from theconventional VH of four chain immunoglobulins. Such a VHH molecule canbe derived from antibodies raised in Camelidae species, for example incamel, llama, dromedary, alpaca and guanaco. Other species besidesCamelidae may produce heavy chain antibodies naturally devoid of lightchain; such VHHs are also contemplated.

The VH and VL regions can be subdivided into regions ofhypervariability, termed “complementarity determining regions” (CDR),interspersed with regions that are more conserved, termed “frameworkregions” (FR). The extent of the framework region and CDRs has beenprecisely defined by a number of methods (see, Kabat, E. A., et al.(1991) Sequences of Proteins of Immunological Interest, Fifth Edition,U.S. Department of Health and Human Services, NIH Publication No.91-3242; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917; and theAbM definition used by Oxford Molecular's AbM antibody modelingsoftware. See, generally, e.g., Protein Sequence and Structure Analysisof Antibody Variable Domains. In: Antibody Engineering Lab Manual (Ed.:Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg). In someembodiments, the following definitions are used: AbM definition of CDR1of the heavy chain variable domain and Kabat definitions for the otherCDRs. In certain embodiments, Kabat definitions are used for all CDRs.In addition, embodiments described with respect to Kabat or AbM CDRs mayalso be implemented using Chothia hypervariable loops. Each VH and VLtypically includes three CDRs and four FRs, arranged from amino-terminusto carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, FR4.

As used herein, an “immunoglobulin variable domain sequence” refers toan amino acid sequence which can form the structure of an immunoglobulinvariable domain. For example, the sequence may include all or part ofthe amino acid sequence of a naturally-occurring variable domain. Forexample, the sequence may or may not include one, two, or more N- orC-terminal amino acids, or may include other alterations that arecompatible with formation of the protein structure.

The term “antigen-binding site” refers to the part of an antibodymolecule that comprises determinants that form an interface that bindsto a TIM-3 polypeptide, or an epitope thereof. With respect to proteins(or protein mimetics), the antigen-binding site typically includes oneor more loops (of at least, e.g., four amino acids or amino acid mimics)that form an interface that binds to the TIM-3 polypeptide. Typically,the antigen-binding site of an antibody molecule includes at least oneor two CDRs, or more typically at least three, four, five or six CDRs.

The terms “compete” or “cross-compete” are used interchangeably hereinto refer to the ability of an antibody molecule to interfere withbinding of an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibodymolecule provided herein, to a target, e.g., human TIM-3. Theinterference with binding can be direct or indirect (e.g., through anallosteric modulation of the antibody molecule or the target). Theextent to which an antibody molecule is able to interfere with thebinding of another antibody molecule to the target, and thereforewhether it can be said to compete, can be determined using a competitionbinding assay, for example, a FACS assay, an ELISA or BIACORE assay. Insome embodiments, a competition binding assay is a quantitativecompetition assay. In some embodiments, a first anti-TIM-3 antibodymolecule is said to compete for binding to the target with a secondanti-TIM-3 antibody molecule when the binding of the first antibodymolecule to the target is reduced by 10% or more, e.g., 20% or more, 30%or more, 40% or more, 50% or more, 55% or more, 60% or more, 65% ormore, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more,95% or more, 98% or more, 99% or more in a competition binding assay(e.g., a competition assay described herein).

As used herein, the term “epitope” refers to the moieties of an antigen(e.g., human TIM-3) that specifically interact with an antibodymolecule. Such moieties, referred to herein as epitopic determinants,typically comprise, or are part of, elements such as amino acid sidechains or sugar side chains. An epitopic determinant can be defined bymethods known in the art or disclosed herein, e.g., by crystallographyor by hydrogen-deuterium exchange. At least one or some of the moietieson the antibody molecule, that specifically interact with an epitopicdeterminant, are typically located in a CDR(s). Typically an epitope hasa specific three dimensional structural characteristics. Typically anepitope has specific charge characteristics. Some epitopes are linearepitopes while others are conformational epitopes.

In an embodiment, an epitopic determinant is a moiety on the antigen,e.g., such as amino acid side chain or sugar side chain, or partthereof, which, when the antigen and antibody molecule areco-crystallized, is within a predetermined distance, e.g., within 5Angstroms, of a moiety on the antibody molecule, referred to herein as a“crystallographic epitopic determinant.” The crystallographic epitopicdeterminants of an epitope are collectively refered to as the“crystallographic epitope.”

A first antibody molecule binds the same epitope as a second antibodymolecule (e.g., a reference antibody molecule, e.g., an antibodymolecule disclosed herein, e.g., ABTIM3-hum21, ABTIM-hum11 orABTIM3-hum03) if the first antibody specifically interacts with the sameepitopic determinants on the antigen as does the second or referenceantibody, e.g., when interaction is measured in the same way for boththe antibody and the second or reference antibody. Epitopes that overlapshare at least one epitopic determinant. A first antibody molecule bindsan overlapping epitope with a second antibody molecule (e.g., areference antibody molecule, e.g., an antibody disclosed herein, e.g.,ABTIM3-hum21, ABTIM-hum11 or ABTIM3-hum03) when both antibody moleculesspecifically interact with a common epitopic determinant. A first and asecond antibody molecule (e.g., a reference antibody molecule, e.g., anantibody molecule disclosed herein, e.g., ABTIM3-hum21, ABTIM-hum11 orABTIM3-hum03) bind substantially overlapping epitopes if at least halfof the epitopic determinants of the second or reference antibody arefound as epitopic determinants in the epitope of the first antibody. Afirst and a second antibody molecule (e.g., a reference antibodymolecule, e.g., an antibody molecule disclosed herein, e.g.,ABTIM3-hum21, ABTIM-hum11 or ABTIM3-hum03) bind substantially the sameepitope if the first antibody molecule binds at least half of the coreepitopic determinants of the epitope of the second or referenceantibody, wherein the core epitopic determinants are defined bycrystallography and hydrogen-deuterium exchange, e.g., includingresidues Val24, Glu25, Thr41, Glu121, Lys122, Phe123, Asn124, Leu125,Lys126, Leu127, Val128, Gly56, Ala57, Cys58, Pro59, Val60, and Phe61 ofhuman TIM-3.

The terms “monoclonal antibody” or “monoclonal antibody composition” asused herein refer to a preparation of antibody molecules of singlemolecular composition. A monoclonal antibody composition displays asingle binding specificity and affinity for a particular epitope. Amonoclonal antibody can be made by hybridoma technology or by methodsthat do not use hybridoma technology (e.g., recombinant methods).

An “effectively human” protein is a protein that does not evoke aneutralizing antibody response, e.g., the human anti-murine antibody(HAMA) response. HAMA can be problematic in a number of circumstances,e.g., if the antibody molecule is administered repeatedly, e.g., intreatment of a chronic or recurrent disease condition. A HAMA responsecan make repeated antibody administration potentially ineffectivebecause of an increased antibody clearance from the serum (see, e.g.,Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)) and alsobecause of potential allergic reactions (see, e.g., LoBuglio et al.,Hybridoma, 5:5117-5123 (1986)).

The antibody molecule can be a polyclonal or a monoclonal antibody. Inother embodiments, the antibody can be recombinantly produced, e.g.,produced by any suitable phage display or combinatorial methods.

Various phage display and combinatorial methods for generatingantibodies are known in the art (as described in, e.g., Ladner et al.U.S. Pat. No. 5,223,409; Kang et al. International Publication No. WO92/18619; Dower et al. International Publication No. WO 91/17271; Winteret al. International Publication WO 92/20791; Markland et al.International Publication No. WO 92/15679; Breitling et al.International Publication WO 93/01288; McCafferty et al. InternationalPublication No. WO 92/01047; Garrard et al. International PublicationNo. WO 92/09690; Ladner et al. International Publication No. WO90/02809; Fuchs et al. (1991) Bio/Technology 9:1370-1372; Hay et al.(1992) Hum Antibod Hybridomas 3:81-85; Huse et al. (1989) Science246:1275-1281; Griffths et al. (1993) EMBO J 12:725-734; Hawkins et al.(1992) J Mol Biol 226:889-896; Clackson et al. (1991) Nature352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991)Bio/Technology 9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res19:4133-4137; and Barbas et al. (1991) PNAS 88:7978-7982, the contentsof all of which are incorporated by reference herein).

In some embodiments, the antibody is a fully human antibody (e.g., anantibody made in a mouse which has been genetically engineered toproduce an antibody from a human immunoglobulin sequence), or anon-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g.,monkey), camel antibody. In certain embodiments, the non-human antibodyis a rodent (mouse or rat antibody). Methods of producing rodentantibodies are known in the art.

Human monoclonal antibodies can be generated using transgenic micecarrying the human immunoglobulin genes rather than the mouse system.Splenocytes from these transgenic mice immunized with the antigen ofinterest are used to produce hybridomas that secrete human mAbs withspecific affinities for epitopes from a human protein (see, e.g., Woodet al. International Application WO 91/00906, Kucherlapati et al. PCTpublication WO 91/10741; Lonberg et al. International Application WO92/03918; Kay et al. International Application 92/03917; Lonberg, N. etal. 1994 Nature 368:856-859; Green, L. L. et al. 1994 Nature Genet.7:13-21; Morrison, S. L. et al. 1994 Proc. Natl. Acad. Sci. USA81:6851-6855; Bruggeman et al. 1993 Year Immunol 7:33-40; Tuaillon etal. 1993 PNAS 90:3720-3724; Bruggeman et al. 1991 Eur J Immunol21:1323-1326).

An antibody can be one in which the variable region, or a portionthereof, e.g., the CDRs, are generated in a non-human organism, e.g., arat or mouse. Chimeric, CDR-grafted, and humanized antibodies are alsocontemplated. Antibodies generated in a non-human organism, e.g., a rator mouse, and then modified, e.g., in the variable framework or constantregion, to decrease antigenicity in a human are also contemplated.

Chimeric antibodies can be produced by any suitable recombinant DNAtechnique. Several are known in the art (see Robinson et al.,International Patent Publication PCT/US86/02269; Akira, et al., EuropeanPatent Application 184,187; Taniguchi, M., European Patent Application171,496; Morrison et al., European Patent Application 173,494; Neubergeret al., International Application WO 86/01533; Cabilly et al. U.S. Pat.No. 4,816,567; Cabilly et al., European Patent Application 125,023;Better et al. (1988 Science 240:1041-1043); Liu et al. (1987) PNAS84:3439-3443; Liu et al., 1987, J. Immunol. 139:3521-3526; Sun et al.(1987) PNAS 84:214-218; Nishimura et al., 1987, Canc. Res. 47:999-1005;Wood et al. (1985) Nature 314:446-449; and Shaw et al., 1988, J. NatlCancer Inst. 80:1553-1559).

A humanized or CDR-grafted antibody will have at least one or two butgenerally all three recipient CDRs (of heavy and or light immunoglobulinchains) replaced with a donor CDR. The antibody may be replaced with atleast a portion of a non-human CDR or only some of the CDRs may bereplaced with non-human CDRs. It is only necessary to replace the numberof CDRs required for binding of the humanized antibody to TIM-3. In someembodiments, the donor will be a rodent antibody, e.g., a rat or mouseantibody, and the recipient will be a human framework or a humanconsensus framework. Typically, the immunoglobulin providing the CDRs iscalled the “donor” and the immunoglobulin providing the framework iscalled the “acceptor.” In some embodiments, the donor immunoglobulin isa non-human (e.g., rodent). The acceptor framework is typically anaturally-occurring (e.g., a human) framework or a consensus framework,or a sequence about 85% or higher, e.g., 90%, 95%, 99% or higheridentical thereto.

As used herein, the term “consensus sequence” refers to the sequenceformed from the most frequently occurring amino acids (or nucleotides)in a family of related sequences (See e.g., Winnaker, From Genes toClones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family ofproteins, each position in the consensus sequence is occupied by theamino acid occurring most frequently at that position in the family. Iftwo amino acids occur equally frequently, either can be included in theconsensus sequence. A “consensus framework” refers to the frameworkregion in the consensus immunoglobulin sequence.

An antibody can be humanized by any suitable method, and several suchmethods known in the art (see e.g., Morrison, S. L., 1985, Science229:1202-1207, by Oi et al., 1986, BioTechniques 4:214, and by Queen etal. U.S. Pat. No. 5,585,089, U.S. Pat. No. 5,693,761 and U.S. Pat. No.5,693,762, the contents of all of which are hereby incorporated byreference).

Humanized or CDR-grafted antibodies can be produced by CDR-grafting orCDR substitution, wherein one, two, or all CDRs of an immunoglobulinchain can be replaced. See e.g., U.S. Pat. No. 5,225,539; Jones et al.1986 Nature 321:552-525; Verhoeyan et al. 1988 Science 239:1534; Beidleret al. 1988 J. Immunol. 141:4053-4060; Winter U.S. Pat. No. 5,225,539,the contents of all of which are hereby expressly incorporated byreference. Winter describes a CDR-grafting method which may be used toprepare humanized antibodies (UK Patent Application GB 2188638A, filedon Mar. 26, 1987; Winter U.S. Pat. No. 5,225,539), the contents of whichis expressly incorporated by reference.

Also provided are humanized antibodies in which specific amino acidshave been substituted, deleted or added. Criteria for selecting aminoacids from the donor are described in, e.g., U.S. Pat. No. 5,585,089,e.g., columns 12-16 of U.S. Pat. No. 5,585,089, the contents of whichare hereby incorporated by reference. Other techniques for humanizingantibodies are described in Padlan et al. EP 519596 A1, published onDec. 23, 1992.

The antibody molecule can be a single chain antibody. A single-chainantibody (scFV) may be engineered (see, for example, Colcher, D. et al.(1999) Ann N Y Acad Sci 880:263-80; and Reiter, Y. (1996) Clin CancerRes 2:245-52). The single chain antibody can be dimerized ormultimerized to generate multivalent antibodies having specificities fordifferent epitopes of the same target protein.

In some embodiments, the antibody molecule has a heavy chain constantregion chosen from, e.g., the heavy chain constant regions of IgG1,IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; particularly, chosenfrom, e.g., the (e.g., human) heavy chain constant regions of IgG1,IgG2, IgG3, and IgG4. In another embodiment, the antibody molecule has alight chain constant region chosen from, e.g., the (e.g., human) lightchain constant regions of kappa or lambda. The constant region can bealtered, e.g., mutated, to modify the properties of the antibody (e.g.,to increase or decrease one or more of: Fc receptor binding, antibodyglycosylation, the number of cysteine residues, effector cell function,and/or complement function). In some embodiments the antibody haseffector function and can fix complement. In other embodiments theantibody does not recruit effector cells or fix complement. In certainembodiments, the antibody has reduced or no ability to bind an Fcreceptor. For example, it may be an isotype or subtype, fragment orother mutant, which does not support binding to an Fc receptor, e.g., ithas a mutagenized or deleted Fc receptor binding region.

The antibody constant region is altered in some embodiments. Methods foraltering an antibody constant region are known in the art. Antibodieswith altered function, e.g. altered affinity for an effector ligand,such as FcR on a cell, or the C1 component of complement can be producedby replacing at least one amino acid residue in the constant portion ofthe antibody with a different residue (see e.g., EP 388,151 A1, U.S.Pat. No. 5,624,821 and U.S. Pat. No. 5,648,260, the contents of all ofwhich are hereby incorporated by reference). Amino acid mutations whichstabilize antibody structure, such as S228P (EU nomenclature, S241P inKabat nomenclature) in human IgG4 are also contemplated. Similar type ofalterations could be described which if applied to the murine, or otherspecies immunoglobulin would reduce or eliminate these functions.

In some embodiments, the only amino acids in the anti-TIM-3 antibodymolecule are canonical amino acids. In some embodiments, the anti-TIM-3antibody molecule comprises naturally-occurring amino acids; analogs,derivatives and congeners thereof; amino acid analogs having variantside chains; and/or all stereoisomers of any of any of the foregoing.The anti-TIM-3 antibody molecule may comprise the D- or L-opticalisomers of amino acids and peptidomimetics.

A polypeptide of an anti-TIM-3 antibody molecule may be linear orbranched, it may comprise modified amino acids, and it may beinterrupted by non-amino acids. The antibody molecule may also bemodified; for example, by disulfide bond formation, glycosylation,lipidation, acetylation, phosphorylation, or any other manipulation,such as conjugation with a labeling component. The polypeptide can beisolated from natural sources, can be a produced by recombinanttechniques from a eukaryotic or prokaryotic host, or can be a product ofsynthetic procedures.

An antibody molecule can be derivatized or linked to another functionalmolecule (e.g., another peptide or protein). As used herein, a“derivatized” antibody molecule is one that has been modified. Methodsof derivatization include but are not limited to the addition of afluorescent moiety, a radionucleotide, a toxin, an enzyme or an affinityligand such as biotin. Accordingly, the antibody molecules are intendedto include derivatized and otherwise modified forms of the antibodiesdescribed herein, including immunoadhesion molecules. For example, anantibody molecule can be functionally linked (by chemical coupling,genetic fusion, noncovalent association or otherwise) to one or moreother molecular entities, such as another antibody (e.g., a bispecificantibody or a diabody), a detectable agent, a cytotoxic agent, apharmaceutical agent, and/or a protein or peptide that can mediateassociation of the antibody or antibody portion with another molecule(such as a streptavidin core region or a polyhistidine tag).

Some types of derivatized antibody molecule are produced by crosslinkingtwo or more antibodies (of the same type or of different types, e.g., tocreate bispecific antibodies). Suitable crosslinkers include those thatare heterobifunctional, having two distinctly reactive groups separatedby an appropriate spacer (e.g., m-maleimidobenzoyl-N-hydroxysuccinimideester) or homobifunctional (e.g., disuccinimidyl suberate). Such linkersare available from Pierce Chemical Company, Rockford, Ill.

Useful detectable agents with which an anti-TIM-3 antibody molecule maybe derivatized (or labeled) to include fluorescent compounds, variousenzymes, prosthetic groups, luminescent materials, bioluminescentmaterials, fluorescent emitting metal atoms, e.g., europium (Eu), andother anthanides, and radioactive materials (described below). Exemplaryfluorescent detectable agents include fluorescein, fluoresceinisothiocyanate, rhodamine, 5dimethylamine-1-napthalenesulfonyl chloride,phycoerythrin and the like. An antibody may also be derivatized withdetectable enzymes, such as alkaline phosphatase, horseradishperoxidase, β-galactosidase, acetylcholinesterase, glucose oxidase andthe like. When an antibody is derivatized with a detectable enzyme, itis detected by adding additional reagents that the enzyme uses toproduce a detectable reaction product. For example, when the detectableagent horseradish peroxidase is present, the addition of hydrogenperoxide and diaminobenzidine leads to a colored reaction product, whichis detectable. An antibody molecule may also be derivatized with aprosthetic group (e.g., streptavidin/biotin and avidin/biotin). Forexample, an antibody may be derivatized with biotin, and detectedthrough indirect measurement of avidin or streptavidin binding. Examplesof suitable fluorescent materials include umbelliferone, fluorescein,fluorescein isothiocyanate, rhodamine, dichlorotriazinylaminefluorescein, dansyl chloride or phycoerythrin; an example of aluminescent material includes luminol; and examples of bioluminescentmaterials include luciferase, luciferin, and aequorin.

Labeled antibody molecule can be used, for example, diagnosticallyand/or experimentally in a number of contexts, including (i) to isolatea predetermined antigen by standard techniques, such as affinitychromatography or immunoprecipitation; (ii) to detect a predeterminedantigen (e.g., in a cellular lysate or cell supernatant) in order toevaluate the abundance and pattern of expression of the protein; (iii)to monitor protein levels in tissue as part of a clinical testingprocedure, e.g., to determine the efficacy of a given treatment regimen.

An antibody molecule may be conjugated to another molecular entity,typically a label or a therapeutic (e.g., immunomodulatory,immunostimularoty, cytotoxic, or cytostatic) agent or moiety.Radioactive isotopes can be used in diagnostic or therapeuticapplications. Radioactive isotopes that can be coupled to the anti-TIM-3antibodies include, but are not limited to α-, β-, or γ-emitters, or β-and γ-emitters. Such radioactive isotopes include, but are not limitedto iodine (¹³¹I or ¹²⁵I), yttrium (⁹⁰ Y), lutetium (¹⁷⁷Lu), actinium(²²⁵Ac), praseodymium, astatine (²¹¹At) rhenium (¹⁸⁶Re), bismuth (²¹²Bior ²¹³Bi), indium (¹¹¹In), technetium (⁹⁹mTc), phosphorus (³²P), rhodium(¹⁸⁸Rh), sulfur (³⁵S), carbon (¹⁴C), tritium (³H), chromium (⁵¹Cr),chlorine (³⁶Cl), cobalt (⁵⁷Co or ⁵⁸Co), iron (⁵⁹Fe), selenium (⁷⁵Se), orgallium (⁶⁷Ga). Radioisotopes useful as therapeutic agents includeyttrium (⁹⁰Y), lutetium (¹⁷⁷Lu), actinium (²²⁵Ac), praseodymium,astatine (²¹¹At), rhenium (¹⁸⁶Re), bismuth (²¹² Bi or ²¹³Bi), andrhodium (¹⁸⁸Rh). Radioisotopes useful as labels, e.g., for use indiagnostics, include iodine (¹³¹I or ¹²⁵I), indium (¹¹¹In), technetium(⁹⁹mTc), phosphorus (³²P), carbon (¹⁴C), and tritium (³ H), or one ormore of the therapeutic isotopes listed above.

The present disclosure provides radiolabeled antibody molecules andmethods of labeling the same. In some embodiments, a method of labelingan antibody molecule is disclosed. The method includes contacting anantibody molecule, with a chelating agent, to thereby produce aconjugated antibody. The conjugated antibody is radiolabeled with aradioisotope, e.g., ¹¹¹Indium, ⁹⁰Yttrium and ¹⁷⁷Lutetium, to therebyproduce a labeled antibody molecule.

As is discussed above, the antibody molecule can be conjugated to atherapeutic agent. Therapeutically active radioisotopes have alreadybeen mentioned. Examples of other therapeutic agents include taxol,cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin,etoposide, tenoposide, vincristine, vinblastine, colchicine,doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone,mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids,procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids,e.g., maytansinol (see U.S. Pat. No. 5,208,020), CC-1065 (see U.S. Pat.Nos. 5,475,092, 5,585,499, 5,846, 545) and analogs or homologs thereof.Therapeutic agents include, but are not limited to, antimetabolites(e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine,5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine,thioepa chlorambucil, CC-1065, melphalan, carmustine (BSNU) andlomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol,streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP)cisplatin), anthracyclinies (e.g., daunorubicin (formerly daunomycin)and doxorubicin), antibiotics (e.g., dactinomycin (formerlyactinomycin), bleomycin, mithramycin, and anthramycin (AMC)), andanti-mitotic agents (e.g., vincristine, vinblastine, taxol andmaytansinoids).

In some aspects, this disclosure provides a method of providing a targetbinding molecule that specifically binds to a TIM-3 receptor. Forexample, the target binding molecule is an antibody molecule. The methodincludes: providing a target protein that comprises at least a portionof non-human protein, the portion being homologous to (at least 70, 75,80, 85, 87, 90, 92, 94, 95, 96, 97, 98, or 99% identical to) acorresponding portion of a human target protein, but differing by atleast one amino acid (e.g., at least one, two, three, four, five, six,seven, eight, or nine amino acids); obtaining an antibody molecule thatspecifically binds to the antigen; and evaluating efficacy of thebinding agent in modulating activity of the target protein. The methodcan further include administering the binding agent (e.g., antibodymolecule) or a derivative (e.g., a humanized antibody molecule) to ahuman subject.

In certain embodiments, the antibody molecule is a multi-specific (e.g.,a bispecific or a trispecific) antibody molecule. Protocols forgenerating bispecific or heterodimeric antibody molecules are known inthe art; including but not limited to, for example, the “knob in a hole”approach described in, e.g., U.S. Pat. No. 5,731,168; the electrostaticsteering Fc pairing as described in, e.g., WO 09/089004, WO 06/106905and WO 2010/129304; Strand Exchange Engineered Domains (SEED)heterodimer formation as described in, e.g., WO 07/110205; Fab armexchange as described in, e.g., WO 08/119353, WO 2011/131746, and WO2013/060867; double antibody conjugate, e.g., by antibody cross-linkingto generate a bi-specific structure using a heterobifunctional reagenthaving an amine-reactive group and a sulfhydryl reactive group asdescribed in, e.g., U.S. Pat. No. 4,433,059; bispecific antibodydeterminants generated by recombining half antibodies (heavy-light chainpairs or Fabs) from different antibodies through cycle of reduction andoxidation of disulfide bonds between the two heavy chains, as describedin, e.g., U.S. Pat. No. 4,444,878; trifunctional antibodies, e.g., threeFab′ fragments cross-linked through sulfhdryl reactive groups, asdescribed in, e.g., U.S. Pat. No. 5,273,743; biosynthetic bindingproteins, e.g., pair of scFvs cross-linked through C-terminal tailspreferably through disulfide or amine-reactive chemical cross-linking,as described in, e.g., U.S. Pat. No. 5,534,254; bifunctional antibodies,e.g., Fab fragments with different binding specificities dimerizedthrough leucine zippers (e.g., c-fos and c-jun) that have replaced theconstant domain, as described in, e.g., U.S. Pat. No. 5,582,996;bispecific and oligospecific mono- and oligovalent receptors, e.g.,VH-CH1 regions of two antibodies (two Fab fragments) linked through apolypeptide spacer between the CH1 region of one antibody and the VHregion of the other antibody typically with associated light chains, asdescribed in, e.g., U.S. Pat. No. 5,591,828; bispecific DNA-antibodyconjugates, e.g., crosslinking of antibodies or Fab fragments through adouble stranded piece of DNA, as described in, e.g., U.S. Pat. No.5,635,602; bispecific fusion proteins, e.g., an expression constructcontaining two scFvs with a hydrophilic helical peptide linker betweenthem and a full constant region, as described in, e.g., U.S. Pat. No.5,637,481; multivalent and multispecific binding proteins, e.g., dimerof polypeptides having first domain with binding region of Ig heavychain variable region, and second domain with binding region of Ig lightchain variable region, generally termed diabodies (higher orderstructures are also encompassed creating for bispecific, trispecific, ortetraspecific molecules, as described in, e.g., U.S. Pat. No. 5,837,242;minibody constructs with linked VL and VH chains further connected withpeptide spacers to an antibody hinge region and CH3 region, which can bedimerized to form bispecific/multivalent molecules, as described in,e.g., U.S. Pat. No. 5,837,821; VH and VL domains linked with a shortpeptide linker (e.g., 5 or 10 amino acids) or no linker at all in eitherorientation, which can form dimers to form bispecific diabodies; trimersand tetramers, as described in, e.g., U.S. Pat. No. 5,844,094; String ofVH domains (or VL domains in family members) connected by peptidelinkages with crosslinkable groups at the C-terminus further associatedwith VL domains to form a series of FVs (or scFvs), as described in,e.g., U.S. Pat. No. 5,864,019; and single chain binding polypeptideswith both a VH and a VL domain linked through a peptide linker arecombined into multivalent structures through non-covalent or chemicalcrosslinking to form, e.g., homobivalent, heterobivalent, trivalent, andtetravalent structures using both scFV or diabody type format, asdescribed in, e.g., U.S. Pat. No. 5,869,620. Additional exemplarymultispecific and bispecific molecules and methods of making the sameare found, for example, in U.S. Pat. Nos. 5,910,573, 5,932,448,5,959,083, 5,989,830, 6,005,079, 6,239,259, 6,294,353, 6,333,396,6,476,198, 6,511,663, 6,670,453, 6,743,896 , 6,809,185, 6,833,441,7,129,330, 7,183,076, 7,521,056, 7,527,787, 7,534,866, 7,612,181,US2002004587A1, US2002076406A1, US2002103345A1, US2003207346A1,US2003211078A1, US2004219643A1, US2004220388A1, US2004242847A1,US2005003403A1, US2005004352A1, US2005069552A1, US2005079170A1,US2005100543A1, US2005136049A1, US2005136051A1, US2005163782A1,US2005266425A1, US2006083747A1, US2006120960A1, US2006204493A1,US2006263367A1, US2007004909A1, US2007087381A1, US2007128150A1,US2007141049A1, US2007154901A1, US2007274985A1, US2008050370A1,US2008069820A1, US2008152645A1, US2008171855A1, US2008241884A1,US2008254512A1, US2008260738A1, US2009130106A1, US2009148905A1,US2009155275A1, US2009162359A1, US2009162360A1, US2009175851A1,US2009175867A1, US2009232811A1, US2009234105A1, US2009263392A1,US2009274649A1, EP346087A2, WO0006605A2, WO02072635A2, WO04081051A1,WO06020258A2, WO2007044887A2, WO2007095338A2, WO2007137760A2,WO2008119353A1, WO2009021754A2, WO2009068630A1, WO9103493A1,WO9323537A1, WO9409131A1, WO9412625A2, WO9509917A1, WO9637621A2,WO9964460A1. The contents of the above-referenced applications areincorporated herein by reference in their entireties.

In other embodiments, the anti-TIM-3 antibody molecule (e.g., amonospecific, bispecific, or multispecific antibody molecule) iscovalently linked, e.g., fused, to another partner e.g., a protein e.g.,one, two or more cytokines, e.g., as a fusion molecule for example afusion protein. In other embodiments, the fusion molecule comprises oneor more proteins, e.g., one, two or more cytokines. In one embodiment,the cytokine is an interleukin (IL) chosen from one, two, three or moreof IL-1, IL-2, IL-12, IL-15 or IL-21. In one embodiment, a bispecificantibody molecule has a first binding specificity to a first target(e.g., to TIM-3), a second binding specificity to a second target (e.g.,LAG-3 or PD-1), and is optionally linked to an interleukin (e.g., IL-12)domain e.g., full length IL-12 or a portion thereof. In otherembodiments, the anti-TIM-3 antibody molecule is fused to anotherprotein e.g., one, two or more cytokines, e.g., as a fusion molecule. Inother embodiments, the fusion molecule comprises one or more proteins,e.g., one, two or more cytokines. In one embodiment, the cytokine is aninterleukin (IL) chosen from one, two, three or more of IL-1, IL-2,IL-12, IL-15 or IL-21.

A “fusion protein” and a “fusion polypeptide” refer to a polypeptidehaving at least two portions covalently linked together, where each ofthe portions is a polypeptide having a different property. The propertymay be a biological property, such as activity in vitro or in vivo. Theproperty can also be simple chemical or physical property, such asbinding to a target molecule, catalysis of a reaction, etc. The twoportions can be linked directly by a single peptide bond or through apeptide linker, but are in reading frame with each other.

Exemplary Anti-TIM-3 Antibody Molecules

In certain embodiments, the anti-TIM-3 antibody comprises:

(a) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQID NO: 10; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and a lightchain variable region (VL) comprising a VLCDR1 amino acid sequence ofSEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and aVLCDR3 amino acid sequence of SEQ ID NO: 14;

(b) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 4; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(c) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 25; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14;

(d) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 24; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8;

(e) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:9; a VHCDR2 amino acid sequence of SEQ ID NO: 31; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14; or

(f) a VH comprising a VHCDR1 amino acid sequence chosen from SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 30; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 9; a VHCDR2 aminoacid sequence of SEQ ID NO: 10; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and a VLCDR3 aminoacid sequence of SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 3; a VHCDR2 aminoacid sequence of SEQ ID NO: 4; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:6, a VLCDR2 amino acid sequence of SEQ ID NO: 7, and a VLCDR3 amino acidsequence of SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 9; a VHCDR2 aminoacid sequence of SEQ ID NO: 25; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and a VLCDR3 aminoacid sequence of SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 3; a VHCDR2 aminoacid sequence of SEQ ID NO: 24; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:6, a VLCDR2 amino acid sequence of SEQ ID NO: 7, and a VLCDR3 amino acidsequence of SEQ ID NO: 8.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 9; a VHCDR2 aminoacid sequence of SEQ ID NO: 31; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and a VLCDR3 aminoacid sequence of SEQ ID NO: 14.

In certain embodiments, the antibody molecule comprises a VH comprisinga VHCDR1 amino acid sequence chosen from SEQ ID NO: 3; a VHCDR2 aminoacid sequence of SEQ ID NO: 30; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:6, a VLCDR2 amino acid sequence of SEQ ID NO: 7, and a VLCDR3 amino acidsequence of SEQ ID NO: 8.

In certain embodiments, the anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 3 or SEQ ID NO: 9; a VHCDR2 amino acidsequence of SEQ ID NO: 4 or SEQ ID NO: 10; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and

(ii) a light chain variable region (VL) comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6 or SEQ ID NO: 12, a VLCDR2 amino acid sequenceof SEQ ID NO: 7 or SEQ ID NO: 13, and a VLCDR3 amino acid sequence ofSEQ ID NO: 8 or SEQ ID NO: 14.

In other embodiments, the anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 3 or SEQ ID NO: 9; a VHCDR2 amino acidsequence of SEQ ID NO: 24 or SEQ ID NO: 25; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and

(ii) a light chain variable region (VL) comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6 or SEQ ID NO: 12, a VLCDR2 amino acid sequenceof SEQ ID NO: 7 or SEQ ID NO: 13, and a VLCDR3 amino acid sequence ofSEQ ID NO: 8 or SEQ ID NO: 14.

In other embodiments, the anti-TIM-3 antibody molecule comprises:

(i) a heavy chain variable region (VH) comprising a VHCDR1 amino acidsequence chosen from SEQ ID NO: 3 or SEQ ID NO: 9; a VHCDR2 amino acidsequence of SEQ ID NO: 30 or SEQ ID NO: 31; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and

(ii) a light chain variable region (VL) comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6 or SEQ ID NO: 12, a VLCDR2 amino acid sequenceof SEQ ID NO: 7 or SEQ ID NO: 13, and a VLCDR3 amino acid sequence ofSEQ ID NO: 8 or SEQ ID NO: 14.

In embodiments of the aforesaid antibody molecules, the VHCDR1 comprisesthe amino acid sequence of SEQ ID NO: 3. In other embodiments, theVHCDR1 comprises the amino acid sequence of SEQ ID NO: 9.

In embodiments of the aforesaid antibody molecules, the VHCDR2 comprisesthe amino acid sequence of SEQ ID NO: 4. In other embodiments, theVHCDR2 comprises the amino acid sequence of SEQ ID NO: 10. In otherembodiments, the VHCDR2 comprises the amino acid sequence of SEQ ID NO:24. In other embodiments, the VHCDR2 comprises the amino acid sequenceof SEQ ID NO: 25. In other embodiments, the VHCDR2 comprises the aminoacid sequence of SEQ ID NO: 30. In other embodiments, the VHCDR2comprises the amino acid sequence of SEQ ID NO: 31.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising an amino acid sequence at least 85%identical to any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68,72, 76, 80, 84, 92, or 100.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:1, 16, 26, 32, 36, 44, 48, 52, 60, 68, 72, 76, 80, 84, 92, or 100.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising an amino acid sequence at least 85%identical to any of SEQ ID NOs: 2, 20, 40, 56, 64, 88, 96, or 104.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:2, 20, 40, 56, 64, 88, 96, or 104.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:1.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:16.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 18.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:26.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 28.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:32.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 34.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:36.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 38.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:44.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 46.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:48.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 50.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:52.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 54.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:60.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 62.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:68.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 70.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:72.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 74.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:76.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 78.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:80.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 82.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:84.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 86.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:92.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 94.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:100.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 102.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 116.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 121.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:2.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:20.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:40.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:56.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:64.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:88.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 90.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:96.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 98.

In other embodiments, the aforesaid antibody molecules comprise a lightchain variable domain comprising the amino acid sequence of SEQ ID NO:104.

In other embodiments, the aforesaid antibody molecules comprise a lightchain comprising the amino acid sequence of SEQ ID NO: 106.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO: 1and a light chain variable domain comprising the amino acid sequence ofSEQ ID NO: 2.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:16 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 20.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:26 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 20.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:32 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 20.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:36 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 40.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:44 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 40.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:48 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 40.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:36 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 20.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:16 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 40.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:52 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:60 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:52 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:60 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:68 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:72 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:76 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:80 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:68 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:72 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 56.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:76 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:80 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 64.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:84 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 88.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:92 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 96.

In other embodiments, the aforesaid antibody molecules comprise a heavychain variable domain comprising the amino acid sequence of SEQ ID NO:100 and a light chain variable domain comprising the amino acid sequenceof SEQ ID NO: 104.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 18 and a lightchain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 28 and a lightchain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 34 and a lightchain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 38 and a lightchain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 46 and a lightchain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 50 and a lightchain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 116 and a lightchain comprising the amino acid sequence of SEQ ID NO: 22.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 121 and a lightchain comprising the amino acid sequence of SEQ ID NO: 42.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 54 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 62 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 54 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 62 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 70 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 74 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 78 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 82 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 70 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 74 and a lightchain comprising the amino acid sequence of SEQ ID NO: 58.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 78 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 82 and a lightchain comprising the amino acid sequence of SEQ ID NO: 66.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 86 and a lightchain comprising the amino acid sequence of SEQ ID NO: 90.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 94 and a lightchain comprising the amino acid sequence of SEQ ID NO: 98.

In other embodiments, the aforesaid antibody molecules comprise a heavychain comprising the amino acid sequence of SEQ ID NO: 102 and a lightchain comprising the amino acid sequence of SEQ ID NO: 106.

In other embodiments, the aforesaid antibody molecules are chosen from aFab, F(ab′)2, Fv, or a single chain Fv fragment (scFv).

In other embodiments, the aforesaid antibody molecules comprise a heavychain constant region selected from IgG1, IgG2, IgG3, and IgG4.

In other embodiments, the aforesaid antibody molecules comprise a lightchain constant region chosen from the light chain constant regions ofkappa or lambda.

In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR2of the VH region of SEQ ID NO: 1, using the Kabat or Chothia definitionsof CDRs. In some embodiments, the anti-TIM-3 antibody molecule comprisesthe CDR2 and one or both of CDR1 and CDR3 of the VH region of SEQ ID NO:1, using the Kabat or Chothia definitions of CDRs. In some embodiments,the anti-TIM-3 antibody molecule comprises CDR2 of the VH region of SEQID NO: 1 in combination with another 1, 2, 3, 4, or 5 (e.g.,collectively all) CDRs found in SEQ ID NO: 1 or SEQ ID NO: 2, using theKabat of Chothia definitions of CDRs. In some embodiments, theanti-TIM-3 antibody molecule comprises the VHCDR2 of SEQ ID NO: 4. Forinstance, the anti-TIM-3 antibody molecule may comprise the VHCDR2 ofSEQ ID NO: 4 in combination with one or both of the VHCDR1 of SEQ ID NO:3 and the VHCDR3 of SEQ ID NO: 5. As a further example, the anti-TIM-3antibody molecule may comprise the VHCDR2 of SEQ ID NO: 4 in combinationwith another 1, 2, 3, 4, or 5 (e.g., collectively all) CDRs selectedfrom SEQ ID NOS: 3, 5, 6, 7, and 8.

In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR3of the VL region of SEQ ID NO: 2, using the Kabat or Chothia definitionsof CDRs. In some embodiments, the anti-TIM-3 antibody molecule comprisesthe CDR3 and one or both of CDR1 and CDR2 of the VL region of SEQ ID NO:2, using the Kabat or Chothia definitions of CDRs. In some embodiments,the anti-TIM-3 antibody molecule comprises CDR3 of the VL region of SEQID NO: 2 in combination with another 1, 2, 3, 4, or 5 (e.g.,collectively all) CDRs found in SEQ ID NO: 1 or SEQ ID NO: 2, using theKabat of Chothia definitions of CDRs. In some embodiments, theanti-TIM-3 antibody molecule comprises the VLCDR3 of SEQ ID NO: 8. Forinstance, the anti-TIM-3 antibody molecule may comprise the VLCDR3 ofSEQ ID NO: 8 in combination with one or both of the VHCDR1 of SEQ ID NO:6 and the VHCDR2 of SEQ ID NO: 7. As a further example, the anti-TIM-3antibody molecule may comprise the VLCDR3 of SEQ ID NO: 8 in combinationwith another 1, 2, 3, 4, or 5 (e.g., collectively all) CDRs selectedfrom SEQ ID NOs: 3-7.

In some embodiments, the anti-TIM-3 antibody molecule comprises the CDR2of the VH region of SEQ ID NO: 1 and the CDR3 of the VL region of SEQ IDNO: 2, optionally in combination with an additional 1, 2, 3, or 4 (e.g.,collectively all) CDRs found in SEQ ID NO: 1 and SEQ ID NO: 2, using theKabat or Chothia definitions of CDRs. In certain embodiments, theanti-TIM-3 antibody molecule comprises the VHCDR2 of SEQ ID NO: 4 andthe VLCDR3 of SEQ ID NO: 8, optionally in combination with an additional1, 2, 3, or 4 (e.g., collectively all) CDRs selected from SEQ ID NOS: 3,5, 6, or 7.

In some embodiments, the anti-TIM-3 antibody molecule comprises a heavychain constant region, a light chain constant region, and heavy andlight chain variable regions of Tables 1-4 (e.g., SEQ ID NO: 1 and SEQID NO: 2). In certain embodiments, the anti-TIM-3 antibody moleculecomprises a heavy chain constant region, a light chain constant region,and 1, 2, 3, 4, 5, or 6 (e.g., all) CDRs of Tables 1-4.

In some embodiments, the anti-TIM-3 antibody molecule comprises thesequence of all or a portion of the heavy chain of SEQ ID NO: 1. Forinstance, in some embodiments, the anti-TIM-3 antibody moleculecomprises amino acids 1-98, 1-107, or 1-118 of SEQ ID NO: 1. In someembodiments, the anti-TIM-3 antibody molecule comprises amino acids 1-98of SEQ ID NO: 1, a hCDR3 region (e.g., SEQ ID NO: 5 or a sequencesubstantially identical thereto), and a VHFW4 region (e.g., a humanVHFW4 region, a homologous region of human D or J sequences, amino acids108-118 of SEQ ID NO: 1, or a sequence substantially identical thereto).In some embodiments, the VHFW4 region has no more than 1 or 2 positionsof non-identity relative to amino acids 108-118 of SEQ ID NO: 1. In someembodiments, the VHFW4 region has no more than 3, 4, 5, 6, 7, 8, 9, or10 positions of non-identity relative to amino acids 108-118 of SEQ IDNO: 1. In some embodiments the hCDR3 region has no more than 1 or 2positions of non-identity relative to SEQ ID NO: 5.

In other embodiments, the aforesaid antibody molecules are capable ofbinding to human TIM-3 with a dissociation constant (K_(D)) of less than0.5 nM.

In some embodiments, the anti-TIM-3 antibody molecule is capable ofindependently binding to human TIM-3 and cynomolgus monkey TIM-3 withhigh affinity. In some embodiments, high affinity refers to a K_(D) ofless than 5, 2, 1, 0.5, 0.4, 0.3, 0.2, or 0.1 nM, e.g., about 0.3 to0.01 nM, e.g., about 0.2 to 0.05 nM, e.g., as measured by a Biacoremethod.

In other embodiments, the aforesaid antibody molecules bind tocynomolgus TIM-3 with a K_(D) of less than 10, 5, 4, 3, 2, or 1 nM,e.g., as measured by a Biacore method, FACS analysis, or ELISA.

In other embodiments, the aforesaid antibody molecules bind to humanTIM-3 with a KD of less than 5, 2, 1, 0.5, 0.4, 0.3, 0.2, or 0.1 nM,e.g., as measured by a Biacore method, FACS analysis, or ELISA.

In embodiments, the aforesaid antibody molecules do not bind to mouseTIM-3.

In some embodiments, the antibody molecule binds to a mammalian, e.g.,human, TIM-3. For example, the antibody molecule binds specifically toan epitope, e.g., linear or conformational epitope, (e.g., an epitope asdescribed herein) on TIM-3. In some embodiments, the epitope is at leasta portion of the IgV domain of human or cynomolgus TIM-3. In certainaspects, it is advantageous to identify an antibody that binds with highaffinity to the human and cynomolgus homologs of a protein of interest.This desirable cross-reactivity allows the same antibody (or twoantibodies with the same CDRs or variable regions) to be tested in ananimal model and then administered to human patients as a therapeutic.

In certain embodiments, the aforesaid antibody molecules are notcross-reactive with mouse TIM-3. In certain embodiments, the aforesaidantibody molecules are less cross-reactive with rat TIM-3. For example,the cross-reactivity can be measured by a Biacore method or a bindingassay using cells that expresses TIM-3 (e.g., human TIM-3-expressing300.19 cells). In other embodiments, the aforesaid antibody moleculesbind an extracellular Ig-like domain (e.g., IgV domain) of TIM-3.

In some embodiments, the aforesaid anti-TIM-3 antibody molecules bind toone or more residues within: the two residues adjacent to the N-terminusof the A strand, the BC loop, the CC′ loop, the F strand, the FG loop,and the G strand of TIM-3, or one or more (e.g., two, five, ten,fifteen, twenty, twenty-five, thirty, thirty-five, or all) residueswithin two or more of the two residues adjacent to the N-terminus of theA strand, the BC loop, the CC′ loop, the F strand, the FG loop, or the Gstrand of TIM-3. The F strand of TIM-3 comprises residues G106 to 1112;the G strand of TIM-3 comprises residues E121 to K130; the FG loop ofTIM-3 comprises the residues between the F strand and the G strand,e.g., comprising residues Q113 to D120; the BC loop of TIM-3 comprisesthe residues between the B strand and the C strand, e.g., comprisingresidues P37 to P50; the two residues adjacent to the N-terminus of theA strand comprises residues V24 and E25; the CC′ loop comprises theresidues between the C strand and the C′ strand, e.g., comprisingresidues G56 to N65. In other embodiments, the aforesaid anti-TIM-3antibody molecules bind to one or more residues within: the A strand,the EF loop, the C strand, the C′C″ loop, or the C″ strand. The A strandcomprises residues Y26 to E29; the EF loop comprises the residuesbetween the E strand and the F strand, e.g., comprising residues E98 toS105; the C strand comprises residues V51 to K55; the C′C″ loopcomprises the residues between the C′ strand and the C″ strand, e.g.,comprising residues D71 to D74; and the C″ strand comprises residues V75to W78. The numbering for the residues of TIM-3 is described, e.g., inFIG. 18. In an embodiment, the anti-TIM-3 antibody molecules bind to oneor more (e.g., two, five, ten, fifteen, twenty, twenty-five, thirty,thirty-five, or all) residues in the F strand, the G strand, and the CC′loop of TIM-3.

In some embodiments, the aforesaid anti-TIM-3 antibody molecules reduceor inhibit plasma membrane penetration or PtdSer-dependent membranepenetration of TIM-3. In some embodiments, the aforesaid anti-TIM-3antibody molecules reduce or inhibit binding to TIM-3 ligand PtdSer. Insome embodiments, the aforesaid anti-TIM-3 antibody molecules reduce orinhibit binding to TIM-3 ligand HMGB1. In some embodiments, theaforesaid anti-TIM-3 antibody molecules reduce or inhibit binding toTIM-3 ligand CEACAM-1. In some embodiments, the aforesaid anti-TIM-3antibody molecules reduce or inhibit binding to TIM-3 ligandSemaphorin-4A. In some embodiments, the aforesaid anti-TIM-3 antibodymolecules do not reduce or inhibit binding to TIM-3 ligand Galectin-9.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, or more continuous or discontinuous (e.g., noncontiguous)amino acid residues chosen from Val24, Glu25, Thr41, Gly56, Ala57,Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124, Leu125,Lys126, and/or Leu127.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous anddiscontinuous (e.g., noncontiguous) amino acid residues chosen fromVal24, Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59,Val60, Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122,Phe123, Asn124, Leu125, Lys126, Leu127, and/or Val128, e.g., as detailedin Table 13.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous ordiscontinuous (e.g., noncontiguous) amino acid residues chosen fromGlu23, Val24, Glu25, Tyr26, Thr41, Pro42, Ala43, Ala44, Pro45, Gly46,Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55, Gly56,Ala57, Cys58, Pro59, Val60, Phe61, Glu121, Lys122, Phe123, Asn124,Leu125, Lys126, and/or Leu127.

In some embodiments, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a TIM-3 surface (e.g., one, two, three, five, eight,ten, fifteen, twenty, twenty-one, twenty-five, or more continuous ordiscontinuous (e.g., noncontiguous) amino acid residues chosen fromVa124, Glu25, Tyr26, Phe39, Tyr40, Thr41, Pro42, Ala43, Ala44, Pro45,Gly46, Asn47, Leu48, Val49, Pro50, Val51, Cys52, Trp53, Gly54, Lys55,Gly56, Ala57, Cys58, Pro59, Val60, Phe61, Ser105, Gly106, Ile107,Asn119, Asp120, Glu121, Lys122, Phe123, Asn124, Leu125, Lys126, Leu127,and/or Val128.

In other embodiments, the anti-TIM-3 antibody molecule competes withCEACAM-1 for binding to TIM-3. In one embodiment, the anti-TIM-3antibody molecule interacts, e.g., binds to, one, two, or more (all) ofC58, N119 and K122 of TIM-3, e.g., displaces or competes CEACAM-1 forbinding to these residues. In one embodiment, the anti-TIM-3 antibodymolecule reduces or blocks the formation of a hydrogen bond between K122of TIM-3 and N42 of CEACAM-1. With respect to CEACAM-1, it has beenshown that CEACAM-1 is a ligand for TIM-3 and is required for itsability to mediate T-cell inhibition, which may have important role inregulating autoimmunity and anti-tumour immunity (Huang, et al. (2014)Nature doi:10.1038/nature13848). Inhibition of an interaction betweenTIM-3 and CEACAM-1 can be used with the other immunomodulators describedherein (e.g., anti-PD-1 inhibitor) to enhance an immune response againsta cancer.

In another embodiment, the anti-TIM-3 antibody molecule interacts with,e.g., binds to, a PtdSer-binding loop of TIM-3, e.g., the human TIM-3IgV domain. In one embodiment, the anti-TIM-3 antibody moleculeinteracts with, e.g., binds to, at least two PtdSer-binding loops ofTIM-3, e.g., the FG loop and CC′ loop of TIM-3 (e.g., a metalion-dependent ligand binding site (MILIBS)). For example, the carboxylgroup of PtdSer can bind to the CC′ loop of TIM-3 and the amino group ofPtdSer can bind to the FG loop of TIM-3. In one embodiment, theanti-TIM-3 antibody molecule reduces or prevents PtdSer-mediatedmembrane penetration of TIM-3 Thus, the anti-TIM-3 antibody molecule mayreduce engagement of TIM-3-expressing cells and/or penetration into themembrane of apoptotic cells (which can display PtdSer) for engulfment.

In another embodiment, the anti-TIM-3 antibody molecule competes withHMGB1 for bind to TIM-3. E.g., it reduces binding of HMGB1 to residue 62of TIM-3 (Q in mouse, E in human TIM-3). With respect to HMGB1, it hasbeen reported to interact with TIM-3 to help tumor-associated dendriticcells suppress nucleic acid-mediated innate immune response (Chiba etal., (2012) Nat. Immunol. 13(9):832-842). Thus, the anti-TIM-3 antibodymolecule may enhance nucleic acid-mediated innate immune response.

In yet another embodiment, the anti-TIM-3 antibody molecule does notcompete with or reduce a Galectin-9 (Gal-9) ligand to binding to TIM-3.

In embodiments, the anti-TIM-3 antibody molecule is a monospecificantibody molecule or a bispecific antibody molecule. In embodiments, theantibody molecule has a first binding specificity for TIM-3 and a secondbinding specifity for PD-1, LAG-3, CEACAM (e.g., CEACAM-1 and/orCEACAM-5), PD-L1 or PD-L2. In embodiments, the antibody moleculecomprises an antigen binding fragment of an antibody, e.g., a halfantibody or antigen binding framgment of a half antibody.

In other embodiments, the aforesaid antibody molecules are capable ofenhancing an antigen-specific T cell response.

Provided herein is an isolated nucleic acid molecule encoding the aboveantibody molecule, vectors and host cells thereof. The nucleic acidmolecule includes but is not limited to RNA, genomic DNA and cDNA.

In embodiments, the isolated nucleic acid encodes the antibody heavychain variable region or light chain variable region, or both, of anythe aforesaid antibody molecules.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a heavy chain variable domain, wherein the nucleotidesequence is at least 85% identical to any of SEQ ID NOs: 11, 17, 29, 33,37, 45, 49, 53, 61, 69, 73, 77, 81, 85, 93, 101, 115, or 120.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a heavy chain variable domain, wherein the nucleotidesequence comprises any of SEQ ID NOs: 11, 17, 27, 33, 37, 45, 49, 53,61, 69, 73, 77, 81, 85, 93, 101, 115, or 120.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a heavy chain, wherein the nucleotide sequence is atleast 85% identical to any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55,63, 71, 75, 79, 83, 87, 95, 103, 117, or 122.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a heavy chain, wherein the nucleotide sequencecomprises any of SEQ ID NOs: 19, 29, 35, 39, 47, 51, 55, 63, 71, 75, 79,83, 87, 95, 103, 117 or 122.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a light chain variable domain, wherein the nucleotidesequence is at least 85% identical to any of SEQ ID NOs: 15, 21, 41, 57,65, 89, 97, 105, 118, 123, 125, or 127.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a light chain variable domain, wherein the nucleotidesequence comprises any of SEQ ID NOs: 15, 21, 41, 57, 65, 89, 97, 105,118, 123, 125, or 127.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a light chain, wherein the nucleotide sequence is atleast 85% identical to any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107,119, 124, 126, or 128.

In other embodiments, the isolated nucleic acid comprises a nucleotidesequence encoding a light chain, wherein the nucleotide sequencecomprises any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107, 119, 124, 126,or 128.

Pharmaceutical Compositions and Kits

In some aspects, this disclosure provides compositions, e.g.,pharmaceutically acceptable compositions, which include an anti-TIM-3antibody molecule described herein, formulated together with apharmaceutically acceptable carrier. As used herein, “pharmaceuticallyacceptable carrier” includes any and all solvents, dispersion media,isotonic and absorption delaying agents, and the like that arephysiologically compatible. The carrier can be suitable for intravenous,intramuscular, subcutaneous, parenteral, rectal, spinal or epidermaladministration (e.g. by injection or infusion).

The compositions set out herein may be in a variety of forms. Theseinclude, for example, liquid, semi-solid and solid dosage forms, such asliquid solutions (e.g., injectable and infusible solutions), dispersionsor suspensions, liposomes, and suppositories. A suitable form depends onthe intended mode of administration and therapeutic application. Typicalsuitable compositions are in the form of injectable or infusiblesolutions. One suitable mode of administration is parenteral (e.g.,intravenous, subcutaneous, intraperitoneal, intramuscular). In someembodiments, the antibody molecule is administered by intravenousinfusion or injection. In certain embodiments, the antibody isadministered by intramuscular or subcutaneous injection.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal, epidural and intrasternal injection andinfusion.

Therapeutic compositions typically should be sterile and stable underthe conditions of manufacture and storage. The composition can beformulated as a solution, microemulsion, dispersion, liposome, or otherordered structure suitable to high antibody concentration. Sterileinjectable solutions can be prepared by incorporating the activecompound (i.e., antibody or antibody portion) in the required amount inan appropriate solvent with one or a combination of ingredientsenumerated above, as required, followed by filtered sterilization.Generally, dispersions are prepared by incorporating the active compoundinto a sterile vehicle that contains a basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum drying and freeze-dryingthat yields a powder of the active ingredient plus any additionaldesired ingredient from a previously sterile-filtered solution thereof.The proper fluidity of a solution can be maintained, for example, by theuse of a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prolonged absorption of injectable compositions can be brought about byincluding in the composition an agent that delays absorption, forexample, monostearate salts and gelatin.

The antibody molecules can be administered by a variety of methods.Several are known in the art, and for many therapeutic applications, anappropriate route/mode of administration is intravenous injection orinfusion. In an embodiment, the antibody molecules can be administeredby intravenous infusion at a rate of more than 20 mg/min, e.g., 20-40mg/min, and preferably greater than or equal to 40 mg/min to reach adose of about 35 to 440 mg/m², preferably about 70 to 310 mg/m², andmore preferably, about 110 to 130 mg/m². In an embodiment, the antibodymolecules can be administered by intravenous infusion at a rate of lessthan 10 mg/min; preferably less than or equal to 5 mg/min to reach adose of about 1 to 100 mg/m², preferably about 5 to 50 mg/m², about 7 to25 mg/m² and more preferably, about 10 mg/m². As will be appreciated bythe skilled artisan, the route and/or mode of administration will varydepending upon the desired results. In certain embodiments, the activecompound may be prepared with a carrier that will protect the compoundagainst rapid release, such as a controlled release formulation,including implants, transdermal patches, and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Many methods for the preparationof such formulations are patented or generally known to those skilled inthe art. See, e.g., Sustained and Controlled Release Drug DeliverySystems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

In certain embodiments, an antibody molecule can be orally administered,for example, with an inert diluent or an assimilable edible carrier. Theantibody molecule (and other ingredients, if desired) may also beenclosed in a hard or soft shell gelatin capsule, compressed intotablets, or incorporated directly into the subject's diet. For oraltherapeutic administration, the antibody molecule may be incorporatedwith excipients and used in the form of ingestible tablets, buccaltablets, troches, capsules, elixirs, suspensions, syrups, wafers, andthe like. To administer an antibody molecule by other than parenteraladministration, it may be necessary to coat the compound with, orco-administer the compound with, a material to prevent its inactivation.Therapeutic compositions can also be administered with medical devices,and several are known in the art.

Dosage regimens are adjusted to provide the desired response (e.g., atherapeutic response). For example, a single bolus may be administered,several divided doses may be administered over time or the dose may beproportionally reduced or increased as indicated by the exigencies ofthe therapeutic situation. It is especially advantageous to formulateparenteral compositions in dosage unit form for ease of administrationand uniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subjects tobe treated; each unit contains a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms are dictated by and directly dependent on (a)the unique characteristics of the active compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding such an active compound for the treatment ofsensitivity in individuals.

An exemplary, non-limiting range for a therapeutically orprophylactically effective amount of an antibody molecule is 0.1-30mg/kg, more preferably 1-25 mg/kg. Dosages and therapeutic regimens ofthe anti-TIM-3 antibody molecule can be determined by a skilled artisan.In certain embodiments, the anti-TIM-3 antibody molecule is administeredby injection (e.g., subcutaneously or intravenously) at a dose of about1 to 40 mg/kg, e.g., 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg, about 10to 20 mg/kg, about 1 to 5 mg/kg, 1 to 10 mg/kg, 5 to 15 mg/kg, 10 to 20mg/kg, 15 to 25 mg/kg, or about 3 mg/kg. The dosing schedule can varyfrom e.g., once a week to once every 2, 3, or 4 weeks. In oneembodiment, the anti-TIM-3 antibody molecule is administered at a dosefrom about 10 to 20 mg/kg every other week. The antibody molecule can beadministered by intravenous infusion at a rate of more than 20 mg/min,e.g., 20-40 mg/min, and preferably greater than or equal to 40 mg/min toreach a dose of about 35 to 440 mg/m², preferably about 70 to 310 mg/m²,and more preferably, about 110 to 130 mg/m². In embodiments, theinfusion rate of about 110 to 130 mg/m² achieves a level of about 3mg/kg. In other embodiments, the antibody molecule can be administeredby intravenous infusion at a rate of less than 10 mg/min, e.g., lessthan or equal to 5 mg/min to reach a dose of about 1 to 100 mg/m², e.g.,about 5 to 50 mg/m², about 7 to 25 mg/m², or, about 10 mg/m². In someembodiments, the antibody is infused over a period of about 30 min. Itis to be noted that dosage values may vary with the type and severity ofthe condition to be alleviated. It is to be further understood that forany particular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions, and that dosage ranges set forth herein are exemplary onlyand are not intended to limit the scope or practice of the claimedcomposition.

The pharmaceutical compositions herein may include a “therapeuticallyeffective amount” or a “prophylactically effective amount” of anantibody molecule. A “therapeutically effective amount” refers to anamount effective, at dosages and for periods of time necessary, toachieve the desired therapeutic result. A therapeutically effectiveamount of the modified antibody or antibody fragment may vary accordingto factors such as the disease state, age, sex, and weight of theindividual, and the ability of the antibody or antibody portion toelicit a desired response in the individual. A therapeutically effectiveamount is also one in which any toxic or detrimental effects of theantibody molecule is outweighed by the therapeutically beneficialeffects. A “therapeutically effective dosage” preferably inhibits ameasurable parameter by at least about 20%, more preferably by at leastabout 40%, even more preferably by at least about 60%, and still morepreferably by at least about 80% relative to untreated subjects. Themeasurable parameter may be, e.g., tumor growth rate or pathogen growthrate. The ability of a compound to inhibit a measurable parameter can beevaluated in an animal model system predictive of efficacy in thecorresponding human disease. Alternatively, this property of acomposition can be evaluated by examining the ability of the compound toinhibit, such inhibition in vitro by assays known to the skilledpractitioner.

A “prophylactically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredprophylactic result. Typically, since a prophylactic dose is used insubjects prior to or at an earlier stage of disease, theprophylactically effective amount will be less than the therapeuticallyeffective amount.

Also within this disclosure is a kit comprising an antibody moleculedescribed herein. The kit can include one or more other elementsincluding: instructions for use; other reagents, e.g., a label, atherapeutic agent, or an agent useful for chelating, or otherwisecoupling, an antibody to a label or therapeutic agent, or aradioprotective composition; devices or other materials for preparingthe antibody molecule for administration; pharmaceutically acceptablecarriers; and devices or other materials for administration to asubject.

Uses of Anti-TIM-3 Antibody Molecules

TIM-3 is a coinhibitory protein expressed, e.g., on activated T helper 1(Th1) CD4+ and cytotoxic CD8+ T cells that secrete IFN-γ. TIM-3 islargely co-expressed on PD-1+ exhausted T cells as shown in preclinicalmodels of cancer and viral exhaustion. Co-blockade of these pathways canrestore effector T cell function (e.g., IFN-γ secretion, proliferation)in several models as well as human PBMCs derived from metastaticmelanoma patients and patients with HIV or HCV. TIM-3 is also enrichedon Fox-P3+ natural regulatory T cells (and FoxP3-negative inducedregulatory cells), and the nTreg expression correlates with diseaseseverity in NSCLC, hepatocellular and ovarian carcinoma. In mousemodels, TIM-3+ nTregs have been shown to be more immunosuppressive(secrete higher levels of IL-10 and TGF-β).

In addition, TIM-3 can play an important role on innate immune cells,including NK cells, monocytes/macrophages and dendritic cells (DCs).TIM-3 is constitutively expressed on macrophages and DCs, and blockadecan enhance TNF-α secretion from human monocytes and increase NF-κBexpression in a mouse dendritic cell line. TIM-3 can also contribute toexpansion of myeloid-derived suppressor cells (MDSCs). Constitutiveexpression of TIM-3 on macrophages is associated with less IL-12secretion, and downregulation of TIM-3 post-TLR activation can lead toenhanced IL-12 and subsequent effector T cell responses.

The antibody molecules disclosed herein have in vitro and in vivodiagnostic, as well as therapeutic and prophylactic utilities. In someembodiments, the antibody molecules modulate (e.g., enhance or inhibit)an immune response in a subject by binding TIM-3. For example, thesemolecules can be administered to cells in culture, in vitro or ex vivo,or to a subject, e.g., a human subject, e.g., in vivo, to modulate(e.g., enhance or inhibit) immunity.

Accordingly, in some aspects, the disclosure provides a method ofmodifying an immune response in a subject comprising administering tothe subject an antibody molecule described herein, such that the immuneresponse in the subject is modified. In some embodiments, the immuneresponse is enhanced, stimulated or up-regulated. In certainembodiments, the immune response is inhibited or downregulated. Forexample, these antibody molecules can be administered to cells inculture, e.g. in vitro or ex vivo, or in a subject, e.g., in vivo, totreat, prevent, and/or diagnose a variety of disorders, such as cancers,immune disorders, and infectious diseases.

As used herein, the term “subject” is intended to include human andnon-human animals. In some embodiments, the subject is a human subject,e.g., a human patient having a disorder or condition characterized byabnormal TIM-3 functioning. Generally, the subject has at least someTIM-3 protein, including the TIM-3 epitope that is bound by the antibodymolecule, e.g., a high enough level of the protein and epitope tosupport antibody binding to TIM-3. The term “non-human animals” includesmammals and non-mammals, such as non-human primates. In someembodiments, the subject is a human. In some embodiments, the subject isa human patient in need of enhancement of an immune response. Themethods and compositions described herein are suitable for treatinghuman patients having a disorder that can be treated by modulating(e.g., augmenting or inhibiting) an immune response.

Methods of Treating Immune Disorders

TIM-3 is a transmembrane receptor expressed on T cells, e.g., CD4+ Tcells, CD8+ T cells, regulatory T cells, and differentiated Th1 cells.TIM-3-dependent trafficking of Th1 cells to target tissue can beinhibited with soluble TIM-3 (see U.S. Pat. No. 7,470,428). Accordingly,modulating TIM-3 function may reduce T-cell trafficking into a targettissue, e.g., in subjects with autoimmune disease. TIM-3 may play animportant role in the induction of autoimmune diseases by regulatingmacrophage activation and/or function. Accordingly, in certainembodiments, the anti-TIM-3 antibody molecules described herein aresuitable for use in downregulating an unwanted immune response, e.g.,treating autoimmune diseases.

Furthermore, as described in the Examples herein, anti-TIM-3 antibodiescan stimulate NK cell-mediated killing of target cells, and can enhanceIFN-gamma secretion and proliferation of CD4+ T cells. Accordingly, incertain embodiments, the anti-TIM-3 antibody molecules described hereinare suitable for use in stimulating a desired immune response, e.g., animmune response against a cancer cell or pathogen.

The anti-TIM-3 antibodies described herein may be used for treatingimmune disorders, especially T lymphocyte-related disorders, including,but not limited to, chronic inflammatory diseases and disorders, such asCrohn's disease, reactive arthritis, including Lyme disease,insulin-dependent diabetes, organ-specific autoimmunity, includingmultiple sclerosis, Hashimoto's thyroiditis and Grave's disease, contactdermatitis, psoriasis, graft rejection, graft versus host disease,sarcoidosis, atopic conditions, such as asthma and allergy, includingallergic rhinitis, gastrointestinal allergies, including food allergies,eosinophilia, conjunctivitis, glomerular nephritis (e.g., IgAnephropathy), certain pathogen susceptibilities such as helminthic(e.g., leishmaniasis).

In certain embodiments, the anti-TIM-3 antibody is used to modulate Tcell function, e.g., CD4+ T cells, CD8+ T cells, Tregs, Th17, and Th1function. In some embodiments, the anti-TIM-3 antibody molecule causesTIM-3 blockade, and is used to treat an immune disorder which is not aTh1-dependent disease (see Schroll et al., Am J Pathol 2010 April;176(4):1716-1742). In certain embodiments, the anti-TIM-3 antibodymolecule does not cause TIM-3 blockade.

In some aspects, the present disclosure provides methods ofadministering an anti-TIM-3 antibody molecule, resulting in promoting orreducing T-cell trafficking to a target tissue, promoting or inhibitingantigen-presenting cell (APC) activation.

In some embodiments the subject is in need of treatment for anautoimmune disease. Autoimmune disease include those in which asubject's own antibodies react with host tissue or in which immuneeffector T cells are autoreactive to endogenous self-peptides and causedestruction of tissue. Thus an immune response is mounted against asubject's own antigens, referred to as self-antigens. Autoimmunediseases include but are not limited to rheumatoid arthritis, Crohn'sdisease e.g., pediatric Crohn's disease, multiple sclerosis, systemiclupus erythematosus (SLE), autoimmune encephalomyelitis, myastheniagravis (MG), Hashimoto's thyroiditis, Goodpasture's syndrome, pemphigus(e.g., pemphigus vulgaris), Grave's disease, autoimmune hemolyticanemia, autoimmune thrombocytopenic purpura, scleroderma withanti-collagen antibodies, mixed connective tissue disease, polymyositis,pernicious anemia, idiopathic Addison's disease, autoimmune-associatedinfertility, glomerulonephritis (e.g., crescentic glomerulonephritis,proliferative glomerulonephritis), bullous pemphigoid, Sjogren'ssyndrome, insulin resistance, autoimmune diabetes mellitus (type 1diabetes mellitus; insulin-dependent diabetes mellitus),atherosclerosis, and Alzheimer's disease.

In some aspects, an anti-TIM-3 antibody molecule described herein isadministered to treat an unwanted immune response to an allergen.Examples of natural animal and plant allergens include proteins specificto the following genuses: Canine (Canis familiaris); Dermatophagoides(e.g., Dermatophagoides farinae); Felis (Felis domesticus); Ambrosia(Ambrosia artemiisfolia; Lolium (e.g., Lolium perenne or Loliummultiflorum); Cryptomeria (Cryptomeria japonica); Alternaria (Alternariaalternata); Alder; Alnus (Alnus gultinosa); Betula (Betula verrucosa);Quercus (Quercus alba); Olea (Olea europa); Artemisia (Artemisiavulgaris); Plantago (e.g., Plantago lanceolata); Parietaria (e.g.,Parietaria officinalis or Parietaria judaica); Blattella (e.g.,Blattella germanica); Apis (e.g., Apis multiflorum); Cupressus (e.g.,Cupressus sempervirens, Cupressus arizonica and Cupressus macrocarpa);Juniperus (e.g., Juniperus sabinoides, Juniperus virginiana, Juniperuscommunis and Juniperus ashei); Thuya (e.g., Thuya orientalis);Chamaecyparis (e.g., Chamaecyparis obtusa); Periplaneta (e.g.,Periplaneta americana); Agropyron (e.g., Agropyron repens); Secale(e.g., Secale cereale); Triticum (e.g., Triticum aestivum); Dactylis(e.g., Dactylis glomerata); Festuca (e.g., Festuca elatior); Poa (e.g.,Poa pratensis or Poa compressa); Avena (e.g., Avena sativa); Holcus(e.g., Holcus lanatus); Anthoxanthum (e.g., Anthoxanthum odoratum);Arrhenatherum (e.g., Arrhenatherum elatius); Agrostis (e.g., Agrostisalba); Phleum (e.g., Phleum pratense); Phalaris (e.g., Phalarisarundinacea); Paspalum (e.g., Paspalum notatum); Sorghum (e.g., Sorghumhalepensis); and Bromus (e.g., Bromus inermis).

In some embodiments, the anti-TIM-3 antibody molecule is administered totreat multiple sclerosis, Crohn's disease, sepsis, SIRS (SystemicInflammatory Response Syndrome), or glomerulonephritis.

Methods of Treating Cancer

In some aspects, the present disclosure provides methods ofadministering an anti-TIM-3 antibody molecule to treat cancer. While notwishing to be bound by theory, in some embodiments, an anti-TIM-3antibody molecule stimulates a patient's immune system to recognize anddestroy cancer cells, thereby treating the cancer. In some embodiments,the cancer to be treated expresses TIM-3, and the anti-TIM-3 antibodymolecule targets the cancer cells or cells in the cancermicroenvironment.

In some aspects, the present disclosure relates to treatment of asubject in vivo using an anti-TIM-3 antibody molecule such that growthof cancerous tumors is inhibited. An anti-TIM-3 antibody may be usedalone to inhibit the growth of cancerous tumors. Alternatively, ananti-TIM-3 antibody may be used in combination with one or more of: astandard cancer treatment (e.g., for cancer or infectious disorders), oranother antibody or antigen-binding fragment thereof, an immunomodulator(e.g., an activator of a costimulatory molecule or an inhibitor of aninhibitory molecule); a vaccine, (e.g., a cancer vaccine); or otherforms of cellular immunotherapy, as described below.

Accordingly, in some embodiments, the disclosure provides a method ofinhibiting growth of tumor cells in a subject, comprising administeringto the subject a therapeutically effective amount of an anti-TIM-3antibody molecule described herein.

In some embodiments, the methods are suitable for the treatment ofcancer in vivo. To achieve antigen-specific enhancement of immunity, theanti-TIM-3 antibody molecule can be administered together with anantigen of interest. When antibodies to TIM-3 are administered incombination with one or more agents, the combination can be administeredin either order or simultaneously.

Types of Cancer

In certain aspects, a method of treating a subject, e.g., reducing orameliorating, a hyperproliferative condition or disorder (e.g., acancer), e.g., solid tumor, a hematological cancer, a soft tissue tumor,or a metastatic lesion, in a subject is provided. The method includesadministering to the subject one or more anti-TIM-3 antibody moleculesdescribed herein, alone or in combination with other agents ortherapeutic modalities.

As used herein, the term “cancer” is meant to include all types ofcancerous growths or oncogenic processes, metastatic tissues ormalignantly transformed cells, tissues, or organs, irrespective ofhistopathologic type or stage of invasiveness. Examples of cancerousdisorders include, but are not limited to, solid tumors, hematologicalcancers, soft tissue tumors, and metastatic lesions. Examples of solidtumors include malignancies, e.g., sarcomas, and carcinomas (includingadenocarcinomas and squamous cell carcinomas) of the various organsystems, such as those affecting liver, lung, breast, lymphoid,gastrointestinal (e.g., colon), genitourinary tract (e.g., renal,urothelial cells), prostate and pharynx. Adenocarcinomas includemalignancies such as most colon cancers, rectal cancer, renal-cellcarcinoma, liver cancer, non-small cell carcinoma of the lung, cancer ofthe small intestine and cancer of the esophagus. Squamous cellcarcinomas include malignancies, e.g., in the lung, esophagus, skin,head and neck region, oral cavity, anus, and cervix. In one embodiment,the cancer is a melanoma, e.g., an advanced stage melanoma. Metastaticlesions of the aforementioned cancers can also be treated or preventedusing the methods and compositions described herein.

Exemplary cancers whose growth can be inhibited using the antibodymolecules disclosed herein include cancers typically responsive toimmunotherapy. Non-limiting examples of suitable cancers for treatmentinclude melanoma (e.g., metastatic malignant melanoma), renal cancer(e.g. clear cell carcinoma), prostate cancer (e.g. hormone refractoryprostate adenocarcinoma), breast cancer, colon cancer and lung cancer(e.g. non-small cell lung cancer). Additionally, refractory or recurrentmalignancies can be treated using the antibody molecules describedherein.

Cancers include, but are not limited to, basal cell carcinoma, biliarytract cancer; bladder cancer; bone cancer; brain and CNS cancer; primaryCNS lymphoma; neoplasm of the central nervous system (CNS); breastcancer; cervical cancer; choriocarcinoma; colon and rectum cancer;connective tissue cancer; cancer of the digestive system; endometrialcancer; esophageal cancer; eye cancer; cancer of the head and neck;gastric cancer; intra-epithelial neoplasm; kidney cancer; larynx cancer;leukemia (including acute myeloid leukemia, chronic myeloid leukemia,acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic oracute leukemias); liver cancer; lung cancer (e.g., small cell andnon-small cell); lymphoma including Hodgkin's and non-Hodgkin'slymphoma; lymphocytic lymphoma; melanoma, e.g., cutaneous or intraocularmalignant melanoma; myeloma; neuroblastoma; oral cavity cancer (e.g.,lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer;prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancerof the respiratory system; sarcoma; skin cancer; stomach cancer;testicular cancer; thyroid cancer; uterine cancer; cancer of the urinarysystem, hepatocarcinoma, cancer of the anal region, carcinoma of thefallopian tubes, carcinoma of the vagina, carcinoma of the vulva, cancerof the small intestine, cancer of the endocrine system, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, solid tumors of childhood,spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi'ssarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma,environmentally induced cancers including those induced by asbestos, aswell as other carcinomas and sarcomas, and combinations of said cancers.

In some embodiments, the cancer treated with the antibody molecules,includes but is not limited to, solid tumors, hematological cancers,soft tissue tumors, and metastatic lesions. Examples of solid tumorsinclude malignancies, e.g., sarcomas, adenocarcinomas, and carcinomas,of the various organ systems, such as those affecting lung, breast,lymphoid, gastrointestinal (e.g., colon), genitals and genitourinarytract (e.g., renal, urothelial, bladder cells), pharynx, CNS (e.g.,brain, neural or glial cells), skin (e.g., melanoma), and pancreas, aswell as adenocarcinomas which include malignancies such as most coloncancers, rectal cancer, renal-cell carcinoma, liver cancer, non-smallcell-carcinoma of the lung, cancer of the small intestine and cancer ofthe esophagus. Methods and compositions disclosed herein are also usefulfor treating metastatic lesions associated with the aforementionedcancers.

While not wishing to be bound by theory, in some embodiments, a patientis more likely to respond to treatment with an immunomodulator(optionally in combination with one or more agents as described herein)if the patient has a cancer that highly expresses PD-L1, and/or thecancer is infiltrated by anti-tumor immune cells, e.g., TILs. Theanti-tumor immunce cells may be positive for CD8, PD-L1, and/or IFN-γ;thus levels of CD8, PD-L1, and/or IFN-γ can serve as a readout forlevels of TILs in the microenvironment. In certain embodiments, thecancer microenvironment is referred to as triple-positive forPD-L1/CD8/IFN-γ.

Accordingly, in certain aspects, this application provides methods ofdetermining whether a tumor sample is positive for one or more of PD-L1,CD8, and IFN-γ, and if the tumor sample is positive for one or more,e.g., two, or all three, of the markers, then administering to thepatient a therapeutically effective amount of an anti-PD-1 antibodymolecule, optionally in combination with one or more otherimmunomodulators or anti-cancer agents, e.g., an anti-TIM3 antibody asdescribed herein.

In the following indications, a large fraction of patients aretriple-positive for PD-L1/CD8/IFN-γ: Lung cancer (squamous); lung cancer(adenocarcinoma); head and neck cancer; stomach cancer; NSCLC; HNSCC;gastric cancers (e.g., MSIhi and/or EBV+); CRC (e.g., MSIhi);nasopharyngeal cancer (NPC); cervical cancer (e.g., squamous); thyroidcancer e.g., papillary thyroid; melanoma; TN breast cancer; and DLBCL(Diffuse Large B-Cell Lymphoma). In breast cancer generally and in coloncancer generally, a moderate fraction of patients is triple-positive forPD-L1/CD8/IFN-γ. In the following indications, a small fraction ofpatients are triple-positive for PD-L1/CD8/IFN-γ: ER+ breast cancer, andpancreatic cancer. These findings are discussed further in Example 9.Regardless of whether a large or small fraction of patients istriple-positive for these markers, screening the patients for thesemarkers allows one to identify a fraction of patients that has anespecially high likelihood of responding favorably to therapy with aPD-1 antibody (e.g., a blocking PD-1 antibody), optionally incombination with one or more other immunomodulators (e.g., an anti-TIM-3antibody molecule described herein, an anti-LAG-3 antibody molecule, oran anti-PD-L1 antibody molecule) and/or anti-cancer agents, e.g., thoselisted in Table 6 and disclosed in the publications listed in Table 6.

In some embodiments, the cancer sample is classified as triple-positivefor PDL1/CD8/IFN-γ. This measurement can roughly be broken down into twothresholds: whether an individual cell is classified as positive, andwhether the sample as a whole is classified as positive. First, one canmeasure, within an individual cell, the level of PD-L1, CD8, and/orIFN-γ. In some embodiments, a cell that is positive for one or more ofthese markers is a cell that has a higher level of the marker comparedto a control cell or a reference value. For example, in someembodiments, a high level of PD-L1 in a given cell is a level higherthan the level of PD-L1 in a corresponding non-cancerous tissue in thepatient. As another example, in some embodiments, a high level of CD8 orIFN-γ in a given cell is a level of that protein typically seen in aTIL. Second, one can also measure the percentage of cells in the samplethat are positive for PD-L1, CD8, and/or IFN-γ. (It is not necessary fora single cell to express all three markers.) In some embodiments, atriple positive sample is one that has a high percentage of cells, e.g.,higher than a reference value or higher than a control sample, that arepositive for these markers.

In other embodiments, one can measure the levels of PDL1, CD8, and/orIFN-γ overall in the sample. In this case, a high level of CD8 or IFN-γin the sample can be the level of that protein typically seen in a tumorinfiltrated with TIL. Similarly, a high level of PD-L1 can be the levelof that protein typically seen in a tumor sample, e.g., a tumormicroenvironment.

The identification of subsets of patients that are triple-positive forPD-L1/CD8/IFN-γ, as shown in Example 10 herein, reveals certainsub-populations of patients that are likely to be especially responsiveto PD-1 antibody therapy. For instance, many IM-TN (immunomodulatory,triple negative) breast cancer patients are triple-positive forPDL1/CD8/IFN-γ. IM-TN breast cancer is described in, e.g., Brian D.Lehmann et al., “Identification of human triple-negative breast cancersubtypes and preclinical models for selection of targeted therapies”, JClin Invest. Jul. 1, 2011; 121(7): 2750-2767. Triple-negative breastcancers are those that do not express estrogen receptor (ER),progesterone receptor (PR) and Her2/neu. These cancers are difficult totreat because they are typically not responsive to agents that targetER, PR, and Her2/neu. Triple-negative breast cancers can be furthersubdivided into different classes, one of which is immunomodulatory. Asdescribed in Lehmann et al., IM-TN breast cancer is enriched for factorsinvolved in immune cell processes, for example, one or more of immunecell signaling (e.g., TH1/TH2 pathway, NK cell pathway, B cell receptorsignaling pathway, DC pathway, and T cell receptor signaling), cytokinesignaling (e.g., cytokine pathway, IL-12 pathway, and IL-7 pathway),antigen processing and presentation, signaling through core immunesignal transduction pathways (e.g., NFKB, TNF, and JAK/STAT signaling),genes involved in T-cell function, immune transcription, interferon(IFN) response and antigen processing. Accordingly, in some embodiments,the cancer treated is a cancer that is, or is determined to be, positivefor one or more marker of IM-TN breast cancer, e.g., a factor thatpromotes one or more of immune cell signaling (e.g., TH1/TH2 pathway, NKcell pathway, B cell receptor signaling pathway, DC pathway, and T cellreceptor signaling), cytokine signaling (e.g., cytokine pathway, IL-12pathway, and IL-7 pathway), antigen processing and presentation,signaling through core immune signal transduction pathways (e.g., NFKB,TNF, and JAK/STAT signaling), genes involved in T-cell function, immunetranscription, interferon (IFN) response and antigen processing.

As another example, it is shown herein that a subset of colon cancerpatients having high MSI (microsatellite instability) is alsotriple-positive for PD-L1/CD8/IFN-γ. Accordingly, in some embodiments, aPD-1 antibody, optionally in combination with one or moreimmunomodulators such as a TIM-3 antibody described herein, a LAG-3antibody, or PD-L1 antibody, and one or more anti-cancer agents, e.g.,an anti-cancer agent described in Table 6 or in a publication in Table6, is administered to a patient who has, or who is identified as having,colon cancer with high MSI, thereby treating the cancer. In someembodiments, a cell with high MSI is a cell having MSI at a level higherthan a reference value or a control cell, e.g., a non-cancerous cell ofthe same tissue type as the cancer.

As another example, it is shown herein that a subset of gastric cancerpatients having high MSI, and/or which is EBV+, is also triple-positivefor PD-L1/CD8/IFN-γ. Accordingly, in some embodiments, a PD-1 antibody,optionally in combination with one or more immunomodulators such as aTIM-3 antibody described herein, a LAG-3 antibody, or PD-L1 antibody,and one or more anti-cancer agents, e.g., an anti-cancer agent describedin Table 6 or in a publication in Table 6 is administered to a patientwho has, or who is identified as having, gastric cancer with high MSIand/or EBV+, thereby treating the cancer. In some embodiments, a cellwith high MSI is a cell having MSI at a level higher than a referencevalue or a control cell, e.g., a non-cancerous cell of the same tissuetype as the cancer.

Additionally disclosed herein are methods of assaying a cancer forPD-L1, and then treating the cancer with a PD-1 antibody, optionally incombination with one or more immunomodulators such as a TIM-3 antibodydescribed herein, a LAG-3 antibody, or PD-L1 antibody. As described inExample 10 herein, a cancer sample can be assayed for PD-L1 proteinlevels or mRNA levels. A sample having levels of PD-L1 (protein or mRNA)higher than a reference value or a control cell (e.g., a non-cancerouscell) can be classified as PD-L1 positive. Accordingly, in someembodiments, a PD-1 antibody (optionally in combination with one or moreanti-cancer agents, optionally in combination with one or moreimmunomodulators such as a TIM-3 antibody described herein, a LAG-3antibody, or PD-L1 antibody) is administered to a patient who has, orwho is identified as having, a cancer that is PD-L1 positive. The cancermay be, e.g., non-small cell lung (NSCLC) adenocarcinoma (ACA), NSCLCsquamous cell carcinoma (SCC), or hepatocellular carcinoma (HCC).

Based on, e.g, Example 9 herein, it was found that certain gastriccancers that are triple-positive for PDL1/CD8/IFN-γ are also positivefor PIK3CA. Accordingly, in some embodiments, a cancer can be treatedwith an anti-PD-1 antibody molecule (optionally in combination with oneor more immunomodulators, e.g., an anti-LAG-3 antibody molecule, ananti-TIM-3 antibody molecule as described herein, or an anti-PD-L1antibody molecule) and an agent that inhibits PIK3CA. Exemplary agentsin this category are described in Stein RC (September 2001). “Prospectsfor phosphoinositide 3-kinase inhibition as a cancer treatment”.Endocrine-related Cancer 8 (3): 237-48 and Marone R, Cmiljanovic V,Giese B, Wymann MP (January 2008). “Targeting phosphoinositide 3-kinase:moving towards therapy”. Biochimica et Biophysica Acta 1784 (1): 159-85.

Based on, e.g, Example 9 herein, CRC, e.g., a patient that has (or isidentified as having) MSI-high CRC may be treated with a PD-1 antibody,optionally in combination with a therapeutic that targets one or more ofTIM-3, e.g., anti-TIM-3 antibody described herein, LAG-3, RNF43, andBRAF. For instance, these cancers may be treated with a PD-1 antibody,optionally in combination with one or more therapeutics that target oneor more of TIM-3, LAG-3, PD-1, RNF43, and BRAF. In embodiments, the oneor more therapeutics include an immunomodulators such as an anti-TIM-3antibody described herein, an anti-LAG-3 antibody molecule, and ananti-cancer agent described in Table 6 or a publication listed in Table6. LAG-3 inhibitors, e.g., antibodies, are described herein. RNF43 canbe inhibited, e.g., with an antibody, small molecule (e.g.,2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28)), siRNA, or a Rspo ligand or derivative thereof. BRAFinhibitors (e.g., vemurafenib or dabrafenib) are described herein.

Based on, e.g, Example 9 herein, a patient that has (or is identified ashaving) a squamous cell lung cancer may be treated with a PD-1 antibodymolecule in combination with a therapeutic that targets TIM-3, e.g., aTIM-3 antibody molecule, LAG-3, e.g., a LAG-3 antibody molecule, andoptionally with one or more anti-cancer agents, e.g., an anti-canceragent described in Table 6 or in a publication in Table 6.

In some embodiments, a subject that has (or is identified as having) asquamous cell lung cancer may be treated with a PD-1 antibody,optionally in combination with a therapeutic that targets TIM-3, e.g., aTIM-3 antibody described herein.

Based on, e.g, Example 9 herein, a patient that has (or is identified ashaving) a thyroid cancer may be treated with a PD-1 antibody molecule,optionally in combination with a therapeutic that targets BRAF, andoptionally in combination with one or more immunomodulators, e.g., ananti-LAG-3 antibody molecule, an anti-TIM-3 antibody molecule describedherein, and an anti-PD-L1 antibody molecule. BRAF inhibitors (e.g.,vemurafenib or dabrafenib) are described herein, e.g., in Table 6 andthe publications listed in Table 6.

In other embodiments, the cancer is a hematological malignancy or cancerincluding but is not limited to a leukemia or a lymphoma. For example, aanti-TIM-3 antibody molecule can be used to treat cancers andmalignancies including, but not limited to, e.g., acute leukemiasincluding but not limited to, e.g., B-cell acute lymphoid leukemia(“BALL”), T-cell acute lymphoid leukemia (“TALL”), acute lymphoidleukemia (ALL); one or more chronic leukemias including but not limitedto, e.g., chronic myelogenous leukemia (CML), chronic lymphocyticleukemia (CLL); additional hematologic cancers or hematologic conditionsincluding, but not limited to, e.g., B cell prolymphocytic leukemia,blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma,diffuse large B cell lymphoma, Follicular lymphoma, Hairy cell leukemia,small cell- or a large cell-follicular lymphoma, malignantlymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma,Marginal zone lymphoma, multiple myeloma, myelodysplasia andmyelodysplastic syndrome, non-Hodgkin's lymphoma, plasmablasticlymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrommacroglobulinemia, and “preleukemia” which are a diverse collection ofhematological conditions united by ineffective production (or dysplasia)of myeloid blood cells, and the like.

In some embodiments, the anti-TIM-3 antibody molecule is used to treat acancer that expresses TIM-3. TIM-3-expressing cancers include cervicalcancer (Cao et al., PLoS One. 2013; 8(1):e53834), lung cancer (Zhuang etal., Am J Clin Pathol. 2012; 137(6):978-985) (e.g., non-small cell lungcancer), acute myeloid leukemia (Kikushige et al., Cell Stem Cell. 2010Dec. 3; 7(6):708-17), diffuse large B cell lymphoma, melanoma (Fourcadeet al., JEM 2010; 207 (10): 2175), renal cancer (e.g., renal cellcarcinoma (RCC), e.g., kidney clear cell carcinoma, kidney papillarycell carcinoma, or metastatic renal cell carcinoma), squamous cellcarcinoma, esophageal squamous cell carcinoma, nasopharyngeal carcinoma,colorectal cancer, breast cancer (e.g., a breast cancer that does notexpress one, two or all of estrogen receptor, progesterone receptor, orHer2/neu, e.g., a triple negative breast cancer), mesothelioma,hepatocellular carcinoma, and ovarian cancer. The TIM-3-expressingcancer may be a metastatic cancer. In other embodiments, the anti-TIM-3antibody molecule is used to treat a cancer that is characterized bymacrophage activity or high expression of macrophage cell markers. In anembodiment, the anti-TIM-3 antibody molecule is used to treat a cancerthat is characterized by high expression of one or more of the followingmacrophage cell markers: LILRB4 (macrophage inhibitory receptor), CD14,CD16, CD68, MSR1, SIGLEC1, TREM2, CD163, ITGAX, ITGAM, CD11b, or CD11c.Examples of such cancers include, but are not limited to, diffuse largeB-cell lymphoma, glioblastoma multiforme, kidney renal clear cellcarcinoma, pancreatic adenocarcinoma, sarcoma, liver heptocellularcarcinoma, lung adenocarcinoma, kidney renal papillary cell carcinoma,skin cutaneous melanoma, brain lower grade glioma, lung squamous cellcarcinoma, ovarian serious cystadenocarcinoma, head and neck squamouscell carcinoma, breast invasive carcinoma, acute myeloid leukemia,cervical squamous cell carcinoma, endocervical adenocarcinoma, uterinecarcinoma, colorectal cancer, uterine corpus endometrial carcinoma,thyroid carcinoma, bladder urothelial carcinoma, adrenocorticalcarcinoma, kidney chromophobe, and prostate adenocarcinoma.

In one embodiment, the cancer is a lung cancer, e.g., a lungadenocarcinoma.

In another embodiment, the cancer is a renal cancer, e.g., a renal cellcarcinoma (RCC) (e.g., a kidney clear cell carcinoma or a kidneypapillary cell carcinoma), or a metastatic lesion thereof.

In yet another embodiment, the cancer is a mesothelioma.

In yet another embodiment, the cancer is a nasopharyngeal carcinoma(NPC).

In yet another embodiment, the cancer is a hematological cancer (e.g., amyeloid leukemia, e.g., acute myeloid leukemia (AML)).

In yet another embodiment, the cancer is a lymphoma (e.g., diffuse largeB cell lymphoma).

In yet another embodiment, the cancer is a breast cancer, e.g., triplenegative (TN) and/or immunomodulatory subtype.

In yet another embodiment, the cancer is glioblastoma multiforme.

In yet another embodiment, the cancer is an ovarian cancer (e.g.,ovarian carcinoma).

In certain embodiments, the cancer is a solid tumor and the antibodymolecule is administered in combination with an anti-LAG-3 or anti-PD-1antibody molecule.

Combination of Anti-TIM-3 Antibodies with Cancer Vaccines

Antibody molecules to TIM-3 can be combined with an immunogenic agent,such as cancerous cells, purified tumor antigens (including recombinantproteins, peptides, and carbohydrate molecules), cells, and cellstransfected with genes encoding immune stimulating cytokines (He et al(2004) J. Immunol. 173:4919-28). Non-limiting examples of tumor vaccinesthat can be used include peptides of melanoma antigens, such as peptidesof gp100, MAGE antigens, Trp-2, MART1 and/or tyrosinase, or tumor cellstransfected to express the cytokine GM-CSF, DNA-based vaccines,RNA-based vaccines, and viral transduction-based vaccines. The cancervaccine may be prophylactic or therapeutic.

In some embodiments, therapy with an anti-TIM-3 antibody molecule iscombined with a vaccination protocol. Many experimental strategies forvaccination against tumors have been devised (see Rosenberg, S., 2000,Development of Cancer Vaccines, ASCO Educational Book Spring: 60-62;Logothetis, C., 2000, ASCO Educational Book Spring: 300-302; Khayat, D.2000, ASCO Educational Book Spring: 414-428; Foon, K. 2000, ASCOEducational Book Spring: 730-738; see also Restifo, N. and Sznol, M.,Cancer Vaccines, Ch. 61, pp. 3023-3043 in DeVita, V. et al. (eds.),1997, Cancer: Principles and Practice of Oncology. Fifth Edition). Inone of these strategies, a vaccine is prepared using autologous orallogeneic tumor cells. These cellular vaccines have been shown to bemost effective when the tumor cells are transduced to express GM-CSF.GM-CSF has been shown to be a potent activator of antigen presentationfor tumor vaccination (Dranoff et al. (1993) Proc. Natl. Acad. Sci.U.S.A. 90: 3539-43).

Anti-TIM-3 antibody molecules can be used in conjunction with acollection of recombinant proteins and/or peptides expressed in a tumorin order to generate an immune response to these proteins. Theseproteins are normally viewed by the immune system as self antigens andare therefore tolerant to them. The tumor antigen may also include theprotein telomerase, which is required for the synthesis of telomeres ofchromosomes and which is expressed in more than 85% of human cancers andin only a limited number of somatic tissues (Kim, N et al. (1994)Science 266: 2011-2013). (These somatic tissues may be protected fromimmune attack by various means). Tumor antigens may also be“neo-antigens” expressed in cancer cells because of somatic mutationsthat alter protein sequence or create fusion proteins between twounrelated sequences (e.g., bcr-abl in the Philadelphia chromosome), oridiotype from B cell tumors.

Other tumor vaccines may include the proteins from viruses implicated inhuman cancers such a Human Papilloma Viruses (HPV), Hepatitis Viruses(HBV and HCV) and Kaposi's Herpes Sarcoma Virus (KHSV), and Epstein-Barrvirus (EBV). Another form of tumor specific antigen which may be used inconjunction with an anti-TIM-3 antibody is purified heat shock proteins(HSP) isolated from the tumor tissue itself. These heat shock proteinscontain fragments of proteins from the tumor cells and these HSPs arehighly efficient at delivery to antigen presenting cells for elicitingtumor immunity (Suot, R & Srivastava, P (1995) Science 269:1585-1588;Tamura, Y. et al. (1997) Science 278:117-120).

Dendritic cells (DC) are potent antigen presenting cells that can beused to prime antigen-specific responses. DC's can be produced ex vivoand loaded with various protein and peptide antigens as well as tumorcell extracts (Nestle, F. et al. (1998) Nature Medicine 4: 328-332). DCsmay also be transduced by genetic means to express these tumor antigensas well. DCs have also been fused directly to tumor cells for thepurposes of immunization (Kugler, A. et al. (2000) Nature Medicine6:332-336). As a method of vaccination, DC immunization may beeffectively combined with an anti-TIM-3 therapy to activate more potentanti-tumor responses.

Alternatively or in combination, the combination further includes aninhibitor or activator of an immune checkpoint modulator, e.g., a LAG-3inhibitor (e.g., an anti-TIM-3 antibody molecule), a PD-L1 inhibitor(e.g., an anti-PD-L1 antibody molecule), a PD-1 inhibitor (e.g., ananti-PD-1 antibody molecule), or a CTLA-4 inhibitor (e.g., ananti-CTLA-4 antibody), or any combination thereof.

TIM-3 blockade may also be combined with a standard cancer treatment.TIM-3 blockade may be effectively combined with chemotherapeuticregimes. In these instances, it may be possible to reduce the dose ofchemotherapeutic reagent administered (Mokyr, M. et al. (1998) CancerResearch 58: 5301-5304). In certain embodiments, the methods andcompositions described herein are administered in combination with oneor more of other antibody molecules, chemotherapy, other anti-cancertherapy (e.g., targeted anti-cancer therapies, or oncolytic drugs),cytotoxic agents, immune-based therapies (e.g., cytokines), surgicaland/or radiation procedures. Exemplary cytotoxic agents that can beadministered in combination with include antimicrotubule agents,topoisomerase inhibitors, anti-metabolites, mitotic inhibitors,alkylating agents, anthracyclines, vinca alkaloids, intercalatingagents, agents capable of interfering with a signal transductionpathway, agents that promote apoptosis, proteosome inhibitors, andradiation (e.g., local or whole body irradiation).

Alternatively, or in combination with the aforesaid combinations, themethods and compositions described herein can be administered incombination with one or more of: an immunomodulator (e.g., an activatorof a costimulatory molecule or an inhibitor of an inhibitory molecule);a vaccine, e.g., a therapeutic cancer vaccine; or other forms ofcellular immunotherapy.

Exemplary non-limiting combinations and uses of the anti-TIM-3 antibodymolecules include the following.

In certain embodiments, the anti-TIM-3 antibody molecule is administeredin combination with a modulator of a costimulatory molecule or aninhibitory molecule, e.g., a co-inhibitory ligand or receptor.

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with a modulator, e.g., agonist, of a costimulatorymolecule. In one embodiment, the agonist of the costimulatory moleculeis chosen from an agonist (e.g., an agonistic antibody orantigen-binding fragment thereof, or soluble fusion) of OX40, CD2, CD27,CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), GITR,CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3or CD83 ligand.

In another embodiment, the anti-TIM-3 antibody molecule is used incombination with a costimulatory molecule, e.g., an agonist associatedwith a positive signal that includes a costimulatory domain of CD28,CD27, ICOS and GITR.

Exemplary GITR agonists include, e.g., GITR fusion proteins andanti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, aGITR fusion protein described in U.S. Pat. No. 6,111,090, EuropeanPatent No.: 090505B1, U.S. Pat. No. 8,586,023, PCT Publication Nos.: WO2010/003118 and 2011/090754, or an anti-GITR antibody described, e.g.,in U.S. Pat. No. 7,025,962, European Patent No.: 1947183B1, U.S. Pat.Nos. 7,812,135, 8,388,967, 8,591,886, European Patent No.: EP 1866339,PCT Publication No.: WO 2011/028683, PCT Publication No.:WO 2013/039954,PCT Publication No.: WO2005/007190, PCT Publication No.: WO 2007/133822,PCT Publication No.: WO2005/055808, PCT Publication No.: WO 99/40196,PCT Publication No.: WO 2001/03720, PCT Publication No.: WO99/20758, PCTPublication No.: WO2006/083289, PCT Publication No.: WO 2005/115451,U.S. Pat. No. 7,618,632, and PCT Publication No.: WO 2011/051726.

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with an inhibitor of an immune checkpoint molecule (orimmune inhibitory molecule). The term “immune checkpoints” as usedherein refers to a group of molecules on the cell surface of immunecells, e.g., CD4 and CD8 T cells that can serve as “brakes” todown-modulate or inhibit an immune response, e.g., an anti-tumor immuneresponse. Immune checkpoint molecules include, but are not limited to,Programmed Death 1 (PD-1), PD-L1, Cytotoxic T-Lymphocyte Antigen 4(CTLA-4), B7-H1, B7-H3, B7-H4, OX-40, 4-1BB (CD137), CD40, T-cellimmunoglobulin domain and mucin domain-3 (TIM-3), andLymphocyte-activation gene 3 (LAG-3), among others. Immunotherapeuticagents that can act as inhibitors of immune checkpoint molecules usefulin combination with the anti-PD-1 molecules described herein, include,but are not limited to, inhibitors of PD-L1, PD-L2, CTLA-4, TIM-3,LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CEACAM (e.g., CEACAM-1,CEACM-3, and/or CEACAM-5), and/or TGFR beta. Inhibition of an immuneinhibitory molecule can be performed by inhibition at the DNA, RNA orprotein level. In embodiments, an inhibitory nucleic acid (e.g., adsRNA, siRNA or shRNA), can be used to inhibit expression of aninhibitory molecule. In other embodiments, the inhibitor of aninhibitory signal is, a polypeptide e.g., a soluble ligand, or anantibody or antigen-binding fragment thereof, that binds to theinhibitory molecule.

In one embodiment, the inhibitor is a soluble ligand (e.g., a CTLA-4-Igor a TIM-3-Ig), or an antibody or antibody fragment that binds toCTLA-4. For example, the anti-TIM-3 antibody molecule can beadministered in combination with an anti-CTLA-4 antibody, e.g.,ipilimumab, for example, to treat a cancer (e.g., a cancer chosen from:a melanoma, e.g., a metastatic melanoma; a lung cancer, e.g., anon-small cell lung carcinoma; or a prostate cancer). Exemplaryanti-CTLA-4 antibodies include Tremelimumab (IgG2 monoclonal antibodyavailable from Pfizer, formerly known as ticilimumab, CP-675,206); andIpilimumab (CTLA-4 antibody, also known as MDX-010, CAS No.477202-00-9). In one embodiment, the anti-TIM-3 antibody molecule isadministered after treatment, e.g., after treatment of a melanoma, withan anti-CTLA-4 antibody (e.g., ipilimumab) with or without a BRAFinhibitor (e.g., vemurafenib or dabrafenib). Exemplary doses that can beuse include a dose of anti-TIM-3 antibody molecule of about 1 to 30mg/kg, 1 to 20 mg/kg, or 1 to 10 mg/kg, e.g., 3 mg/kg, and a dose of ananti-CTLA-4 antibody, e.g., ipilimumab, of about 3 mg/kg.

In certain embodiments, immune checkpoint molecules, e.g., PD-1, LAG-3,TIM-3, CEACAM-1/-5, can regulate T-cell function to promote tumoralimmune escape. Thus, the anti-TIM-3 antibodies described herein can beused in combination with one or more inhibitors of these immuneinhibitor molecules to enhance an anti-tumor response. The combinationof antibodies recited herein can be administered separately, e.g., asseparate antibodies, or linked, e.g., as a bispecific or trispecificantibody molecule.

In one embodiment, the anti-TIM-3 antibody molecule is administered incombination with an anti-TIM-3 antibody or an antigen-binding fragmentthereof. In another embodiment, the anti-TIM-3 antibody molecule isadministered in combination with an anti-PD-1 antibody orantigen-binding fragment thereof. In yet other embodiments, theanti-TIM-3 antibody molecule is administered in combination with ananti-TIM-3 antibody and an anti-PD-1 antibody, or antigen-bindingfragments thereof. In one embodiment, a bispecific antibody thatincludes an anti-TIM-3 antibody molecule and an anti-PD-1 or anti-TIM-3antibody, or antigen-binding fragment thereof, is administered. Incertain embodiments, the combination of antibodies recited herein isused to treat a cancer, e.g., a cancer as described herein (e.g., asolid tumor). The efficacy of the aforesaid combinations can be testedin animal models known in the art. For example, the animal models totest the effect of anti-PD-1 and anti-LAG-3 are described, e.g., in Wooet al. (2012) Cancer Res. 72(4):917-27).

In some embodiments, the inhibitors of the TIM-3 and PD-1 molecules(e.g., anti-TIM-3 and anti-PD-1 antibody molecules) are administered incombination, e.g., to treat cancer. In some embodiments, the subject isa patient who has progressed (e.g., experienced tumor growth) duringtherapy with a PD-1 inhibitor (e.g., an antibody molecule as describedherein) and/or a PD-L1 inhibitor (e.g., an anti-PD-L1 antibodymolecule). In some embodiments, therapy with the PD-1 antibody moleculeand/or PD-L1 antibody molecule is continued, and a TIM-3 immuneinhibitory molecule (e.g., antibody) is added to the therapy.

In other embodiments, the anti-TIM-3 antibody molecule is administeredin combination with a CEACAM inhibitor (e.g., CEACAM-1, CEACAM-3, and/orCEACAM-5 inhibitor). In one embodiment, the inhibitor of CEACAM is ananti-CEACAM antibody molecule. In one embodiment, the anti-TIM-3antibody molecule is administered in combination with a CEACAM-1inhibitor, e.g., an anti-CEACAM-1 antibody molecule. In anotherembodiment, the anti-TIM-3 antibody molecule is administered incombination with a CEACAM-3 inhibitor, e.g., an anti-CEACAM-3 antibodymolecule. In another embodiment, the anti-TIM-3 antibody molecule isadministered in combination with a CEACAM-5 inhibitor, e.g., ananti-CEACAM-5 antibody molecule. Exemplary anti-CEACAM-1 antibodies aredescribed in WO 2010/125571, WO 2013/082366 and WO 2014/022332, e.g., amonoclonal antibody 34B1, 26H7, and 5F4; or a recombinant form thereof,as described in, e.g., US 2004/0047858, U.S. Pat. No. 7,132,255 and WO99/052552. In other embodiments, the anti-CEACAM antibody binds toCEACAM-5 as described in, e.g., Zheng et al. PLoS One. 2010 Sep. 2;5(9). pii: e12529 (DOI:10:1371/journal.pone.0021146), or crossreactswith CEACAM-1 and CEACAM-5 as described in, e.g., WO 2013/054331 and US2014/0271618.

Without wishing to be bound by theory, carcinoembryonic antigen celladhesion molecules (CEACAM), such as CEACAM-1 and CEACAM-5, are believedto mediate, at least in part, inhibition of an anti-tumor immuneresponse (see e.g., Markel et al. J Immunol. 2002 Mar. 15;168(6):2803-10; Markel et al. J Immunol. 2006 Nov. 1; 177(9):6062-71;Markel et al. Immunology. 2009 February; 126(2):186-200; Markel et al.Cancer Immunol Immunother. 2010 February; 59(2):215-30; Ortenberg et al.Mol Cancer Ther. 2012 June; 11(6):1300-10; Stern et al. J Immunol. 2005Jun. 1; 174(11):6692-701; Zheng et al. PLoS One. 2010 Sep. 2; 5(9). pii:e12529). For example, CEACAM-1 has been described as a heterophilicligand for TIM-3 and as playing a role in TIM-3-mediated T celltolerance and exhaustion (see e.g., WO 2014/022332; Huang, et al. (2014)Nature doi:10.1038/nature13848). In embodiments, co-blockade of CEACAM-1and TIM-3 has been shown to enhance an anti-tumor immune response inxenograft colorectal cancer models (see e.g., WO 2014/022332; Huang, etal. (2014), supra). In other embodiments, co-blockade of CEACAM-1 andPD-1 reduce T cell tolerance as described, e.g., in WO 2014/059251.Thus, CEACAM inhibitors can be used with the other immunomodulatorsdescribed herein (e.g., anti-PD-1 and/or anti-TIM-3 inhibitors) toenhance an immune response against a cancer, e.g., a melanoma, a lungcancer (e.g., NSCLC), a bladder cancer, a colon cancer an ovariancancer, and other cancers as described herein.

In some embodiments, the PD-1 and TIM-3 immune inhibitory molecules(e.g., antibody molecules) are administered in combination with eachother, e.g., to treat cancer. In some embodiments, the patient is apatient who progressed (e.g., experienced tumor growth) during therapywith a PD-1 inhibitor (e.g., an antibody molecule as described herein)and/or a PDL1 inhibitor (e.g., antibody molecule). In some embodiments,therapy with the PD-1 antibody molecule and/or PDL1 antibody molecule iscontinued, and a TIM-3 immune inhibitory molecule (e.g., antibody) isadded to the therapy.

In some embodiments, the TIM-3 and LAG-3 immune inhibitory molecules(e.g., antibody molecules) are administered in combination with eachother, e.g., to treat cancer. In some embodiments, the patient is apatient who progressed (e.g., experienced tumor growth) during therapywith a TIM-3 inhibitor (e.g., an antibody molecule as described herein)and/or a PD-1 inhibitor (e.g., antibody molecule). In some embodiments,therapy with the anti-TIM-3 antibody molecule and/or PDL1 antibodymolecule is continued, and a LAG-3 immune inhibitory molecule (e.g.,antibody) is added to the therapy.

In other embodiments, the anti-TIM-3 antibody molecule is administeredin combination with a cytokine, e.g., interleukin-21, interleukin-2, orinterleukin 15. In certain embodiments, the combination of anti-TIM-3antibody molecule and cytokine described herein is used to treat acancer, e.g., a cancer as described herein (e.g., a solid tumor ormelanoma).

Exemplary immunomodulators that can be used in combination withanti-TIM-3 antibody molecules include, but are not limited to, e.g.,afutuzumab (available from Roche®); pegfilgrastim (Neulasta®);lenalidomide (CC-5013, Revlimid®); thalidomide (Thalomid®), actimid(CC4047); and cytokines, e.g., IL-21 or IRX-2 (mixture of humancytokines including interleukin 1, interleukin 2, and interferon γ, CAS951209-71-5, available from IRX Therapeutics).

In yet other embodiments, the anti-TIM-3 antibody molecule is used incombination with an indolearnine-pyrrole 2,3-dioxygenase (IDO) inhibitor(e.g., INCB24360) in a subject with advanced or metastatic cancer (e.g.,a patient with metastic and recurrent NSCL cancer).

In other embodiments, the anti-TIM-3 antibody molecules are administeredto a subject in conjunction with (e.g., before, simultaneously orfollowing) one or more of: bone marrow transplantation, T cell ablativetherapy using chemotherapy agents such as, fludarabine, external-beamradiation therapy (XRT), cyclophosphamide, and/or antibodies such asOKT3 or CAMPATH. In one embodiment, the anti-TIM-3 antibody moleculesare administered following B-cell ablative therapy such as agents thatreact with CD20, e.g., Rituxan. For example, in one embodiment, subjectsmay undergo standard treatment with high dose chemotherapy followed byperipheral blood stem cell transplantation. In certain embodiments,following the transplant, subjects receive the anti-TIM-3 antibodymolecules. In an additional embodiment, the anti-TIM-3 antibodymolecules are administered before or following surgery.

Another example of a combination is an anti-TIM-3 antibody incombination with decarbazine for the treatment of melanoma. Withoutbeing bound by theory, the combined use of TIM-3 blockade andchemotherapy is believed to be facilitated by cell death, that is aconsequence of the cytotoxic action of most chemotherapeutic compounds,which can result in increased levels of tumor antigen in the antigenpresentation pathway. Other combination therapies that may result insynergy with TIM-3 blockade through cell death are radiation, surgery,and hormone deprivation. Each of these protocols creates a source oftumor antigen in the host. Angiogenesis inhibitors may also be combinedwith TIM-3 blockade. Inhibition of angiogenesis leads to tumor celldeath which may feed tumor antigen into host antigen presentationpathways.

TIM-3 blocking antibodies can also be used in combination withbispecific antibodies. Bispecific antibodies can be used to target twoseparate antigens. For example anti-Fc receptor/anti tumor antigen(e.g., Her-2/neu) bispecific antibodies have been used to targetmacrophages to sites of tumor. This targeting may more effectivelyactivate tumor specific responses. The T cell arm of these responseswould be augmented by the use of TIM-3 blockade. Alternatively, antigenmay be delivered directly to DCs by the use of bispecific antibodieswhich bind to tumor antigen and a dendritic cell specific cell surfacemarker.

Tumors evade host immune surveillance by a large variety of mechanisms.Many of these mechanisms may be overcome by the inactivation of proteinswhich are expressed by the tumors and which are immunosuppressive. Theseinclude among others TGF-beta (Kehrl, J. et al. (1986) J. Exp. Med. 163:1037-1050), IL-10 (Howard, M. & O'Garra, A. (1992) Immunology Today 13:198-200), and Fas ligand (Hahne, M. et al. (1996) Science 274:1363-1365). Antibodies or antigen-binding fragments thereof to each ofthese entities may be used in combination with anti-TIM-3 antibodymolecules to counteract the effects of the immunosuppressive agent andfavor tumor immune responses by the host.

Other antibodies which may be used to activate host immuneresponsiveness can be used in combination with anti-TIM-3 antibodymolecules. These include molecules on the surface of dendritic cellswhich activate DC function and antigen presentation. Anti-CD40antibodies are able to substitute effectively for T cell helper activity(Ridge, J. et al. (1998) Nature 393: 474-478) and can be used inconjunction with PD-1 antibodies (Ito, N. et al. (2000) Immunobiology201 (5) 527-40). Antibodies to T cell costimulatory molecules such asCTLA-4 (e.g., U.S. Pat. No. 5,811,097), OX-40 (Weinberg, A. et al.(2000) Immunol 164: 2160-2169), 4-1BB (Melero, I. et al. (1997) NatureMedicine 3: 682-685 (1997), and ICOS (Hutloff, A. et al. (1999) Nature397: 262-266) may also provide for increased levels of T cellactivation.

Additional exemplary standard of care treatments are described in thesection entitled “Combination Therapies” below.

In all of the methods described herein, TIM-3 blockade can be combinedwith other forms of immunotherapy such as cytokine treatment (e.g.,interferons, GM-CSF, G-CSF, IL-2, IL-21), or bispecific antibodytherapy, which provides for enhanced presentation of tumor antigens (seee.g., Holliger (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak(1994) Structure 2:1121-1123).

Methods of administering the antibody molecules are known in the art andare described below. Suitable dosages of the molecules used will dependon the age and weight of the subject and the particular drug used.Dosages and therapeutic regimens of the anti-TIM-3 antibody molecule canbe determined by a skilled artisan. In certain embodiments, theanti-TIM-3 antibody molecule is administered by injection (e.g.,subcutaneously or intravenously) at a dose of about 1 to 30 mg/kg, e.g.,about 5 to 25 mg/kg, about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about3 mg/kg. In some embodiments, the anti-TIM-3 antibody molecule isadministered at a dose of about 1 mg/kg, about 3 mg/kg, about 10 mg/kg,about 15 mg/kg, about 20 mg/kg, about 25 mg/kg or about 30 mg/kg. Insome embodiments, the anti-TIM-3 antibody molecule is administered at adose of about 1-3 mg/kg, about 3-10 mg/kg, about 3-15 mg/kg, about 10-15mg/kg, about 10-20 mg/kg, about 10-25 mg/kg, or about 20-30 mg/kg. Insome embodiments, the anti-TIM-3 antibody molecule is administered at adose of about 0.5-2, 2-4, 2-5, or 5-15 mg/kg. The dosing schedule canvary from e.g., once a week to once every 2, 3, or 4 weeks. In oneembodiment, the anti-TIM-3 antibody molecule is administered at a dosefrom about 10 to 20 mg/kg every other week.

The antibody molecules can be used by themselves or conjugated to asecond agent, e.g., a cytotoxic drug, radioisotope, or a protein, e.g.,a protein toxin or a viral protein. This method includes: administeringthe antibody molecule, alone or conjugated to a cytotoxic drug, to asubject requiring such treatment. The antibody molecules can be used todeliver a variety of therapeutic agents, e.g., a cytotoxic moiety, e.g.,a therapeutic drug, a radioisotope, molecules of plant, fungal, orbacterial origin, or biological proteins (e.g., protein toxins) orparticles (e.g., a recombinant viral particles, e.g.; via a viral coatprotein), or mixtures thereof.

Anti-TIM-3 antibody molecules may also be combined with standard cancertreatments. For instance, anti-TIM-3 antibody molecules may beeffectively combined with chemotherapeutic regimes. In these instances,it may be possible to reduce the dose of chemotherapeutic reagentadministered (Mokyr, M. et al. (1998) Cancer Research 58: 5301-5304). Anexample of such a combination is an anti-TIM-3 antibody molecule incombination with decarbazine for the treatment of melanoma. Anotherexample of such a combination is an anti-TIM-3 antibody molecule incombination with interleukin-2 (IL-2) for the treatment of melanoma. Insome embodiments the anti-TIM-3 antibody molecule can be combined withIL-21. While not wishing to be bound by theory, one scientific rationalebehind the combined use of anti-TIM-3 antibody molecule therapy andchemotherapy is that cell death, that is a consequence of the cytotoxicaction of most chemotherapeutic compounds, should result in increasedlevels of tumor antigen in the antigen presentation pathway. Othercombination therapies that may result in synergy with anti-TIM-3antibody molecule therapy through cell death are radiation, surgery, andhormone deprivation. Each of these protocols creates a source of tumorantigen in the host. Angiogenesis inhibitors may also be combined withanti-TIM-3 antibody molecule therapy. Inhibition of angiogenesis leadsto tumor cell death which may feed tumor antigen into host antigenpresentation pathways. Anti-TIM-3 antibody molecules can also be used incombination with bispecific antibodies. Bispecific antibodies can beused to target two separate antigens. For example anti-Fc receptor/antitumor antigen (e.g., Her-2/neu) bispecific antibodies have been used totarget macrophages to sites of tumor. This targeting may moreeffectively activate tumor specific responses. The T cell arm of theseresponses would be augmented by the use of anti-TIM-3 antibodymolecules. Alternatively, antigen may be delivered directly to DCs bythe use of bispecific antibodies which bind to tumor antigen and adendritic cell specific cell surface marker.

Tumors evade host immune surveillance by a large variety of mechanisms.Many of these mechanisms may be overcome by the inactivation of proteinswhich are expressed by the tumors and which are immunosuppressive. Theseinclude among others TGF-beta (Kehrl, J. et al. (1986) J. Exp. Med. 163:1037-1050), IL-10 (Howard, M. & O'Garra, A. (1992) Immunology Today 13:198-200), and Fas ligand (Hahne, M. et al. (1996) Science 274:1363-1365). Antibodies to each of these entities may be used incombination with anti-TIM-3 antibody molecules to counteract the effectsof the immunosuppressive agent and favor tumor immune responses by thehost.

Other antibodies which may be used to activate host immuneresponsiveness can be used in combination with anti-TIM-3 antibodymolecules. These include molecules on the surface of dendritic cellswhich activate DC function and antigen presentation. Anti-CD40antibodies are able to substitute effectively for T cell helper activity(Ridge, J. et al. (1998) Nature 393: 474-478) and can be used inconjunction with anti-TIM-3 antibody molecules (see Ito, N. et al.(2000) Immunobiology 201 (5) 527-40). Activating antibodies to T cellcostimulatory molecules such as CTLA-4 (e.g., U.S. Pat. No. 5,811,097),OX-40 (Weinberg, A. et al. (2000) Immunol 164: 2160-2169), 4-1BB(Melero, I. et al. (1997) Nature Medicine 3: 682-685 (1997), and ICOS(Hutloff, A. et al. (1999) Nature 397: 262-266) may also provide forincreased levels of T cell activation.

Additional Combination Therapies

The anti-TIM-3 antibody molecule can be used in combination with othertherapies. For example, the combination therapy can include acomposition of the present invention co-formulated with, and/orco-administered with, one or more additional therapeutic agents, e.g.,one or more anti-cancer agents, cytotoxic or cytostatic agents, hormonetreatment, vaccines, and/or other immunotherapies. In other embodiments,the antibody molecules are administered in combination with othertherapeutic treatment modalities, including surgery, radiation,cryosurgery, and/or thermotherapy. Such combination therapies mayadvantageously utilize lower dosages of the administered therapeuticagents, thus avoiding possible toxicities or complications associatedwith the various monotherapies.

By “in combination with,” it is not intended to imply that the therapyor the therapeutic agents must be administered at the same time and/orformulated for delivery together, although these methods of delivery arewithin the scope described herein. The anti-TIM-3 antibody molecules canbe administered concurrently with, prior to, or subsequent to, one ormore other additional therapies or therapeutic agents. The anti-TIM-3antibody molecule and the other agent or therapeutic protocol can beadministered in any order. In general, each agent will be administeredat a dose and/or on a time schedule determined for that agent. In willfurther be appreciated that the additional therapeutic agent utilized inthis combination may be administered together in a single composition oradministered separately in different compositions. In general, it isexpected that additional therapeutic agents utilized in combination beutilized at levels that do not exceed the levels at which they areutilized individually. In some embodiments, the levels utilized incombination will be lower than those utilized individually.

In certain embodiments, the anti-TIM-3 antibody molecules describedherein are administered in combination with one or more other inhibitorsof TIM-3 or other immune checkpoint molecules, e.g., PD-1, PD-L1, PD-L2,CEACAM (e.g., CEACAM-1, CEACAM-3, or CEACAM-5), or LAG-3.

In certain embodiments, the anti-TIM-3 antibody molecules describedherein are administered in combination with one or more other inhibitorsof PD-1, PD-L1 and/or PD-L2 known in the art. The antagonist may be anantibody, an antigen binding fragment thereof, an immunoadhesin, afusion protein, or oligopeptide. In some embodiments, the anti-PD-1antibody is chosen from MDX-1106, Merck 3475 or CT-011. In someembodiments, the PD-1 inhibitor is an immunoadhesin (e.g., animmunoadhesin comprising an extracellular or PD-1 binding portion ofPD-L1 or PD-L2 fused to a constant region (e.g., an Fc region of animmunoglobulin sequence). In some embodiments, the PD-1 inhibitor isAMP-224. In some embodiments, the PD-L1 inhibitor is anti-PD-L1antibody. In some embodiments, the anti-PD-L1 binding antagonist ischosen from YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C, orMDX-1105. MDX-1105, also known as BMS-936559, is an anti-PD-L1 antibodydescribed in WO2007/005874. Antibody YW243.55.S70 (heavy and light chainvariable region sequences shown in SEQ ID Nos. 20 and 21, respectively)is an anti-PD-L1 described in WO 2010/077634.

MDX-1106, also known as MDX-1106-04, ONO-4538 or BMS-936558, is ananti-PD-1 antibody described in WO2006/121168. Merck 3745, also known asMK-3475 or SCH-900475, is an anti-PD-1 antibody described inWO2009/114335. Pidilizumab (CT-011; Cure Tech) is a humanized IgG1kmonoclonal antibody that binds to PD-1. Pidilizumab and other humanizedanti-PD-1 monoclonal antibodies are disclosed in WO2009/101611. In otherembodiments, the anti-PD-1 antibody is pembrolizumab. Pembrolizumab(Trade name Keytruda formerly lambrolizumab-also known as MK-3475)disclosed, e.g., in Hamid, O. et al. (2013) New England Journal ofMedicine 369 (2): 134-44. AMP-224 (B7-DCIg; Amplimmune; e.g., disclosedin WO2010/027827 and WO2011/066342), is a PD-L2 Fc fusion solublereceptor that blocks the interaction between PD-1 and B7-H1. Otheranti-PD-1 antibodies include AMP 514 (Amplimmune), among others, e.g.,anti-PD-1 antibodies disclosed in U.S. Pat. No. 8,609,089, US2010028330, and/or US 20120114649.

In some embodiments, the anti-PD-1 antibody is MDX-1106. Alternativenames for MDX-1106 include MDX-1106-04, ONO-4538, BMS-936558 orNivolumab. In some embodiments, the anti-PD-1 antibody is Nivolumab (CASRegistry Number: 946414-94-4). Nivolumab (also referred to as BMS-936558or MDX1106; Bristol-Myers Squibb) is a fully human IgG4 monoclonalantibody which specifically blocks PD-1. Nivolumab (clone 5C4) and otherhuman monoclonal antibodies that specifically bind to PD-1 are disclosedin U.S. Pat. No. 8,008,449 and WO2006/121168. Pembrolizumab (Trade nameKeytruda formerly lambrolizumab also known as MK-3475; Merck) is ahumanized IgG4 monoclonal antibody that binds to PD-1. Lambrolizumab andother humanized anti-PD-1 antibodies are disclosed in U.S. Pat. No.8,354,509 and WO2009/114335. MDPL3280A (Genentech/Roche) is a human Fcoptimized IgG1 monoclonal antibody that binds to PD-L1. MDPL3280A andother human monoclonal antibodies to PD-L1 are disclosed in U.S. Pat.No. 7,943,743 and U.S Publication No.: 20120039906. Other anti-PD-L1binding agents include YW243.55.570 (heavy and light chain variableregions are shown in SEQ ID NOs 20 and 21 in WO2010/077634) and MDX-1105(also referred to as BMS-936559, and, e.g., anti-PD-L1 binding agentsdisclosed in WO2007/005874).

Cancer Therapies

Exemplary combinations of anti-TIM-3 antibody molecules (alone or incombination with other stimulatory agents) and standard of care forcancer, include at least the following. In certain embodiments, theanti-TIM-3 antibody molecule, e.g., the anti-TIM-3 antibody moleculedescribed herein, is used in combination with a standard of cancer carechemotherapeutic agent including, but not limited to, anastrozole(Arimidex®), bicalutamide (Casodex®), bleomycin sulfate (Blenoxane®),busulfan (Myleran®), busulfan injection (Busulfex®), capecitabine(Xeloda®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin(Paraplatin®), carmustine (BiCNU®), chlorambucil (Leukeran®), cisplatin(Platinol®), cladribine (Leustatin®), cyclophosphamide (Cytoxan® orNeosar®), cytarabine, cytosine arabinoside (Cytosar-U®), cytarabineliposome injection (DepoCyt®), dacarbazine (DTIC-Dome®), dactinomycin(Actinomycin D, Cosmegan), daunorubicin hydrochloride (Cerubidine®),daunorubicin citrate liposome injection (DaunoXome®), dexamethasone,docetaxel (Taxotere®), doxorubicin hydrochloride (Adriamycin®, Rubex®),etoposide (Vepesid®), fludarabine phosphate (Fludara®), 5-fluorouracil(Adrucil®, Efudex®), flutamide (Eulexin®), tezacitibine, Gemcitabine(difluorodeoxycitidine), hydroxyurea (Hydrea®), Idarubicin (Idamycin®),ifosfamide (IFEX®), irinotecan (Camptosar®), L-asparaginase (ELSPAR®),leucovorin calcium, melphalan (Alkeran®), 6-mercaptopurine(Purinethol®), methotrexate (Folex®), mitoxantrone (Novantrone®),mylotarg, paclitaxel (Taxol®), phoenix (Yttrium90/MX-DTPA), pentostatin,polifeprosan 20 with carmustine implant (Gliadel®), tamoxifen citrate(Nolvadex®), teniposide (Vumon®), 6-thioguanine, thiotepa, tirapazamine(Tirazone®), topotecan hydrochloride for injection (Hycamptin®),vinblastine (Velban®), vincristine (Oncovin®), and vinorelbine(Navelbine®), Ibrutinib, idelalisib, and brentuximab vedotin.

Exemplary alkylating agents include, without limitation, nitrogenmustards, ethylenimine derivatives, alkyl sulfonates, nitrosoureas andtriazenes): uracil mustard (Aminouracil Mustard®, Chlorethaminacil®,Demethyldopan®, Desmethyldopan®, Haemanthamine®, Nordopan®, Uracilnitrogen Mustard®, Uracillost®, Uracilmostaza®, Uramustin®,Uramustine®), chlormethine (Mustargen®), cyclophosphamide (Cytoxan®,Neosar®, Clafen®, Endoxan®, Procytox®, Revimmune™), ifosfamide(Mitoxana®), melphalan (Alkeran®), Chlorambucil (Leukeran®), pipobroman(Amedel®, Vercyte®), triethylenemelamine (Hemel®, Hexalen®, Hexastat®),triethylenethiophosphoramine, Temozolomide (Temodar®), thiotepa(Thioplex®), busulfan (Busilvex®, Myleran®), carmustine (BiCNU®),lomustine (CeeNU®), streptozocin (Zanosar®), and Dacarbazine(DTIC-Dome®). Additional exemplary alkylating agents include, withoutlimitation, Oxaliplatin (Eloxatin®); Temozolomide (Temodar® andTemodal®); Dactinomycin (also known as actinomycin-D, Cosmegen®);Melphalan (also known as L-PAM, L-sarcolysin, and phenylalanine mustard,Alkeran®); Altretamine (also known as hexamethylmelamine (HMM),Hexalen®); Carmustine (BiCNU®); Bendamustine (Treanda®); Busulfan(Busulfex® and Myleran®); Carboplatin (Paraplatin®); Lomustine (alsoknown as CCNU, CeeNU®); Cisplatin (also known as CDDP, Platinol® andPlatinol®-AQ); Chlorambucil (Leukeran®); Cyclophosphamide (Cytoxan® andNeosar®); Dacarbazine (also known as DTIC, DIC and imidazolecarboxamide, DTIC-Dome®); Altretamine (also known as hexamethylmelamine(HMM), Hexalen®); Ifosfamide (Ifex®); Prednumustine; Procarbazine(Matulane®); Mechlorethamine (also known as nitrogen mustard, mustineand mechloroethamine hydrochloride, Mustargen®); Streptozocin(Zanosar®); Thiotepa (also known as thiophosphoamide, TESPA and TSPA,Thioplex®); Cyclophosphamide (Endoxan®, Cytoxan®, Neosar®, Procytox®,Revimmune®); and Bendamustine HCl (Treanda®).

Exemplary anthracyclines include, e.g., doxorubicin (Adriamycin® andRubex®); bleomycin (Lenoxane®); daunorubicin (dauorubicin hydrochloride,daunomycin, and rubidomycin hydrochloride, Cerubidine®); daunorubicinliposomal (daunorubicin citrate liposome, DaunoXome®); mitoxantrone(DHAD, Novantrone®); epirubicin (Ellence™); idarubicin (Idamycin®,Idamycin PFS®); mitomycin C (Mutamycin®); geldanamycin; herbimycin;ravidomycin; and desacetylravidomycin.

Exemplary vinca alkaloids that can be used in combination with theanti-PD-1 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-TIM-3 antibodymolecule), include, but ate not limited to, vinorelbine tartrate(Navelbine®), Vincristine (Oncovin®), and Vindesine (Eldisine®));vinblastine (also known as vinblastine sulfate, vincaleukoblastine andVLB, Alkaban-AQ® and Velban®); and vinorelbine (Navelbine®).

Exemplary proteosome inhibitors that can be used in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), include, but ate not limited to, bortezomib (Velcade®);carfilzomib (PX-171-007,(S)-4-Methyl-N-((S)-1-(((S)-4-methyl-1-((R)-2-methyloxiran-2-yl)-1-oxopentan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)-2-((S)-2-(2-morpholinoacetamido)-4-phenylbutanamido)-pentanamide);marizomib (NPI-0052); ixazomib citrate (MLN-9708); delanzomib(CEP-18770); andO-Methyl-N-[(2-methyl-5-thiazolyl)carbonyl]-L-seryl-O-methyl-N-[(1S)-2-[(2R)-2-methyl-2-oxiranyl]-2-oxo-1-(phenylmethyl)ethyl]-L-serinamide(ONX-0912).

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a tyrosine kinaseinhibitor (e.g., a receptor tyrosine kinase (RTK) inhibitor). Exemplarytyrosine kinase inhibitor include, but are not limited to, an epidermalgrowth factor (EGF) pathway inhibitor (e.g., an epidermal growth factorreceptor (EGFR) inhibitor), a vascular endothelial growth factor (VEGF)pathway inhibitor (e.g., a vascular endothelial growth factor receptor(VEGFR) inhibitor (e.g., a VEGFR-1 inhibitor, a VEGFR-2 inhibitor, aVEGFR-3 inhibitor)), a platelet derived growth factor (PDGF) pathwayinhibitor (e.g., a platelet derived growth factor receptor (PDGFR)inhibitor (e.g., a PDGFR-β inhibitor)), a RAF-1 inhibitor, a KITinhibitor and a RET inhibitor. In some embodiments, the anti-canceragent used in combination with the hedgehog inhibitor is selected fromthe group consisting of: axitinib (AG013736), bosutinib (SKI-606),cediranib (RECENTIN™, AZD2171), dasatinib (SPRYCEL®, BMS-354825),erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®,CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®), lestaurtinib(CEP-701), neratinib (HKI-272), nilotinib (TASIGNA®), semaxanib(semaxinib, SU5416), sunitinib (SUTENT®, SU11248), toceranib(PALLADIA®), vandetanib (ZACTIMA®, ZD6474), vatalanib (PTK787, PTK/ZK),trastuzumab (HERCEPTIN®), bevacizumab (AVASTIN®), rituximab (RITUXAN®),cetuximab (ERBITUX®), panitumumab (VECTIBIX®), ranibizumab (Lucentis®),nilotinib (TASIGNA®), sorafenib (NEXAVAR®), alemtuzumab (CAMPATH®),gemtuzumab ozogamicin (MYLOTARG®), ENMD-2076, PCI-32765, AC220,dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523,PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154,CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, XL228,AEE788, AG-490, AST-6, BMS-599626, CUDC-101, PD153035, pelitinib(EKB-569), vandetanib (zactima), WZ3146, WZ4002, WZ8040, ABT-869(linifanib), AEE788, AP24534 (ponatinib), AV-951(tivozanib), axitinib,BAY 73-4506 (regorafenib), brivanib alaninate (BMS-582664), brivanib(BMS-540215), cediranib (AZD2171), CHIR-258 (dovitinib), CP 673451,CYC116, E7080, Ki8751, masitinib (AB1010), MGCD-265, motesanibdiphosphate (AMG-706), MP-470, OSI-930, Pazopanib Hydrochloride,PD173074, Sorafenib Tosylate(Bay 43-9006), SU 5402, TSU-68(SU6668),vatalanib, XL880 (GSK1363089, EXEL-2880). Selected tyrosine kinaseinhibitors are chosen from sunitinib, erlotinib, gefitinib, orsorafenib.

In certain embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a VascularEndothelial Growth Factor (VEGF) receptor inhibitors, including but notlimited to, Bevacizumab (Avastin®), axitinib (Inlyta®); Brivanibalaninate (BMS-582664,(S)-((R)-1-(4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-5-methylpyrrolo[2,1-f][1,2,4]triazin-6-yloxy)propan-2-yl)2-aminopropanoate);Sorafenib (Nexavar®); Pazopanib (Votrient®); Sunitinib malate (Sutent®);Cediranib (AZD2171, CAS 288383-20-1); Vargatef (BIBF1120, CAS928326-83-4); Foretinib (GSK1363089); Telatinib (BAY57-9352, CAS332012-40-5); Apatinib (YN968D1, CAS 811803-05-1); Imatinib (Gleevec®);Ponatinib (AP24534, CAS 943319-70-8); Tivozanib (AV951, CAS475108-18-0); Regorafenib (BAY73-4506, CAS 755037-03-7); Vatalanibdihydrochloride (PTK787, CAS 212141-51-0); Brivanib (BMS-540215, CAS649735-46-6); Vandetanib (Caprelsa® or AZD6474); Motesanib diphosphate(AMG706, CAS 857876-30-3,N-(2,3-dihydro-3,3-dimethyl-1H-indol-6-yl)-2-[(4-pyridinylmethyl)amino]-3-pyridinecarboxamide,described in PCT Publication No. WO 02/066470); Dovitinib dilactic acid(TKI258, CAS 852433-84-2); Linfanib (ABT869, CAS 796967-16-3);Cabozantinib (XL184, CAS 849217-68-1); Lestaurtinib (CAS 111358-88-4);N-[5-[[[5-(1,1-Dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide(BMS38703, CAS 345627-80-7);(3R,4R)-4-Amino-1-((4-((3-methoxyphenyl)amino)pyrrolo[2,1-f][1,2,4]triazin-5-yl)methyl)piperidin-3-ol(BMS690514);N-(3,4-Dichloro-2-fluorophenyl)-6-methoxy-7-[[(3aα,5β,6aα)-octahydro-2-methylcyclopenta[c]pyrrol-5-yl]methoxy]-4-quinazolinamine(XL647, CAS 781613-23-8);4-Methyl-3-[[1-methyl-6-(3-pyridinyl)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]amino]-N-[3-(trifluoromethyl)phenyl]-benzamide(BHG712, CAS 940310-85-0); and Aflibercept (Eylea®).

Exemplary anti-VEGF antibodies include, but are not limited to, amonoclonal antibody that binds to the same epitope as the monoclonalanti-VEGF antibody A4.6.1 produced by hybridoma ATCC HB 10709; arecombinant humanized anti-VEGF monoclonal antibody generated accordingto Presta et al. (1997) Cancer Res. 57:4593-4599. In one embodiment, theanti-VEGF antibody is Bevacizumab (BV), also known as rhuMAb VEGF orAVASTIN®. It comprises mutated human IgG1 framework regions andantigen-binding complementarity-determining regions from the murineanti-hVEGF monoclonal antibody A.4.6.1 that blocks binding of human VEGFto its receptors. Bevacizumab and other humanized anti-VEGF antibodiesare further described in U.S. Pat. No. 6,884,879 issued Feb. 26, 2005.Additional antibodies include the G6 or B20 series antibodies (e.g.,G6-31, B20-4.1), as described in PCT Publication No. WO2005/012359, PCTPublication No. WO2005/044853, the contents of these patent applicationsare expressly incorporated herein by reference. For additionalantibodies see U.S. Pat. Nos. 7,060,269, 6,582,959, 6,703,020,6,054,297, WO98/45332, WO 96/30046, WO94/10202, EP 0666868B1, U.S.Patent Application Publication Nos. 2006009360, 20050186208,20030206899, 20030190317, 20030203409, and 20050112126; and Popkov etal, Journal of Immunological Methods 288: 149-164 (2004). Otherantibodies include those that bind to a functional epitope on human VEGFcomprising of residues F17, M1 8, D19, Y21, Y25, Q89, 191, K1 01, E1 03,and C104 or, alternatively, comprising residues F17, Y21, Q22, Y25, D63,183 and Q89.

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a PI3K inhibitor.In one embodiment, the PI3K inhibitor is an inhibitor of delta and gammaisoforms of PI3K. Exemplary PI3K inhibitors that can be used incombination are described in, e.g., WO 2010/036380, WO 2010/006086, WO09/114870, WO 05/113556, GSK 2126458, GDC-0980, GDC-0941, Sanofi XL147,XL756, XL147, PF-46915032, BKM 120, CAL-101, CAL 263, SF1126, PX-886,and a dual PI3K inhibitor (e.g., Novartis BEZ235).

In some embodiments, the anti-TIM-3 antibody molecules described hereinis used, alone or in combination with another immunomodulator (e.g., ananti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), in combinationwith a mTOR inhibitor, e.g., one or more mTOR inhibitors chosen from oneor more of rapamycin, temsirolimus (TORISEL®), AZD8055, BEZ235, BGT226,XL765, PF-4691502, GDC0980, SF1126, OSI-027, GSK1059615, KU-0063794,WYE-354, Palomid 529 (P529), PF-04691502, or PKI-587. ridaforolimus(formally known as deferolimus, (1R,2R,4S)-4-[(2R)-2[(1R,9S,12S,15R,16E,18R,19R,21R,23S,24E,26E,28Z,30S,32S,35R)-1,18-dihydroxy-19,30-dimethoxy-15,17,21,23,29,35-hexamethyl-2,3,10,14,20-pentaoxo-11,36-dioxa-4-azatricyclo[30.3.1.0^(4,9)]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]-2-methoxycyclohexyldimethylphosphinate, also known as AP23573 and MK8669, and described inPCT Publication No. WO 03/064383); everolimus (Afinitor® or RAD001);rapamycin (AY22989, Sirolimus®); simapimod (CAS 164301-51-3);emsirolimus,(5-{2,4-Bis[(3S)-3-methylmorpholin-4-yl]pyrido[2,3-d]pyrimidin-7-yl}-2-methoxyphenyl)methanol(AZD8055);2-Amino-8-[trans-4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxy-3-pyridinyl)-4-methyl-pyrido[2,3-d]pyrimidin-7(8H)-one(PF04691502, CAS 1013101-36-4); andN²-[1,4-dioxo-4-[[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholinium-4-yl]methoxy]butyl]-L-arginylglycyl-L-α-aspartylL-serine-,inner salt (SF1126, CAS 936487-67-1), and XL765.

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a BRAF inhibitor,e.g., GSK2118436, RG7204, PLX4032, GDC-0879, PLX4720, and sorafenibtosylate (Bay 43-9006).

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a MEK inhibitor.In some embodiments, the combination of the anti-TIM-3 antibody and theMEK inhibitor is used to treat a cancer (e.g., a cancer describedherein). In some embodiments, the cancer treated with the combination ischosen from a melanoma, a colorectal cancer, a non-small cell lungcancer, an ovarian cancer, a breast cancer, a prostate cancer, apancreatic cancer, a hematological malignancy or a renal cell carcinoma.In certain embodiments, the cancer includes a BRAF mutation (e.g., aBRAF V600E mutation), a BRAF wildtype, a KRAS wildtype or an activatingKRAS mutation. The cancer may be at an early, intermediate or latestage. Any MEK inhibitor can be used in combination including, but notlimited to, ARRY-142886, G02442104 (also known as GSK1120212), RDEA436,RDEA119/BAY 869766, AS703026, G00039805 (also known as AZD-6244 orselumetinib), BIX 02188, BIX 02189, CI-1040 (PD-184352), PD0325901,PD98059, U0126, GDC-0973 (Methanone,[3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl][3-hydroxy-3-(25)-2-piperidinyl-1-azetidinyl]-),G-38963, G02443714 (also known as AS703206), or a pharmaceuticallyacceptable salt or solvate thereof. Additional examples of MEKinhibitors are disclosed in WO 2013/019906, WO 03/077914, WO2005/121142, WO 2007/04415, WO 2008/024725 and WO 2009/085983, thecontents of which are incorporated herein by reference.

In some embodiments, the anti-TIM-3 antibody molecule, e.g., theanti-TIM-3 antibody molecule described herein, is used, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), in combination with a JAK2 inhibitor,e.g., CEP-701, INCB18424, CP-690550 (tasocitinib).

In some embodiments, the pharmaceutical composition described herein isused, alone or in combination with another immunomodulator (e.g., ananti-LAG-3, anti-PD-L1 or anti-PD-1 antibody molecule), in combinationwith paclitaxel or a paclitaxel agent, e.g., TAXOL®, protein-boundpaclitaxel (e.g., ABRAXANE®). Exemplary paclitaxel agents include, butare not limited to, nanoparticle albumin-bound paclitaxel (ABRAXANE,marketed by Abraxis Bioscience), docosahexaenoic acid bound-paclitaxel(DHA-paclitaxel, Taxoprexin, marketed by Protarga), polyglutamatebound-paclitaxel (PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX,marketed by Cell Therapeutic), the tumor-activated prodrug (TAP), ANG105(Angiopep-2 bound to three molecules of paclitaxel, marketed byImmunoGen), paclitaxel-EC-1 (paclitaxel bound to the erbB2-recognizingpeptide EC-1; see Li et al., Biopolymers (2007) 87:225-230), andglucose-conjugated paclitaxel (e.g., 2′-paclitaxel methyl2-glucopyranosyl succinate, see Liu et al., Bioorganic & MedicinalChemistry Letters (2007) 17:617-620).

Radiation therapy can be administered through one of several methods, ora combination of methods, including without limitation external-beamtherapy, internal radiation therapy, implant radiation, stereotacticradiosurgery, systemic radiation therapy, radiotherapy and permanent ortemporary interstitial brachytherapy. The term “brachytherapy,” refersto radiation therapy delivered by a spatially confined radioactivematerial inserted into the body at or near a tumor or otherproliferative tissue disease site. The term is intended withoutlimitation to include exposure to radioactive isotopes (e.g. At-211,I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, andradioactive isotopes of Lu). Suitable radiation sources for use as acell conditioner of the present invention include both solids andliquids. By way of non-limiting example, the radiation source can be aradionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source,I-125 as a solid source, or other radionuclides that emit photons, betaparticles, gamma radiation, or other therapeutic rays. The radioactivematerial can also be a fluid made from any solution of radionuclide(s),e.g., a solution of I-125 or I-131, or a radioactive fluid can beproduced using a slurry of a suitable fluid containing small particlesof solid radionuclides, such as Au-198, Y-90. Moreover, theradionuclide(s) can be embodied in a gel or radioactive micro spheres.

Anti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibodymolecule), can be administered in combination with one or more of theexisting modalities for treating cancers, including, but not limited to:surgery; radiation therapy (e.g., external-beam therapy which involvesthree dimensional, conformal radiation therapy where the field ofradiation is designed, local radiation (e.g., radition directed to apreselected target or organ), or focused radiation). Focused radiationcan be selected from the group consisting of stereotactic radiosurgery,fractionated stereotactic radiosurgery, and intensity-modulatedradiation therapy. The focused radiation can have a radiation sourceselected from the group consisting of a particle beam (proton),cobalt-60 (photon), and a linear accelerator (x-ray), e.g., as decribedin WO 2012/177624.

In certain embodiments, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is administered in combination with anantibody against a Killer-cell Immunoglobulin-like Receptors (alsoreferred to herein as an “anti-KIR antibody”), a pan-KIR antibody, or ananti-NKG2D antibody, and an anti-MICA antibody. In certain embodiments,the combination of anti-TIM-3 antibody molecule and anti-KIR antibody,pan-KIR antibody, or an anti-NKG2D antibody described herein is used totreat a cancer, e.g., a cancer as described herein (e.g., a solid tumor,e.g., an advanced solid tumor).

In one embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is administered in combination with acellular immunotherapy (e.g., Provenge (e.g., Sipuleucel)), andoptionally in combination with cyclophosphamide. In certain embodiments,the combination of anti-TIM-3 antibody molecule, Provenge and/orcyclophosphamide is used to treat a cancer, e.g., a cancer as describedherein (e.g., a prostate cancer, e.g., an advanced prostate cancer).

In another embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is administered in combination with avaccine, e.g., a dendritic cell renal carcinoma (DC-RCC) vaccine. Incertain embodiments, the combination of anti-TIM-3 antibody molecule andthe DC-RCC vaccine is used to treat a cancer, e.g., a cancer asdescribed herein (e.g., a renal carcinoma, e.g., metastatic renal cellcarcinoma (RCC) or clear cell renal cell carcinoma (CCRCC)).

In yet another embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is administered in combination withchemotherapy, and/or immunotherapy. For example, the anti-TIM-3 antibodymolecule can be used to treat a myeloma, alone or in combination withone or more of: chemotherapy or other anti-cancer agents (e.g.,thalidomide analogs, e.g., lenalidomide), an anti-PD-1 antibody, tumorantigen-pulsed dendritic cells, fusions (e.g., electrofusions) of tumorcells and dendritic cells, or vaccination with immunoglobulin idiotypeproduced by malignant plasma cells. In one embodiment, the anti-TIM-3antibody molecule is used in combination with an anti-PD-1 antibody totreat a myeloma, e.g., a multiple myeloma.

In one embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is used in combination withchemotherapy to treat a lung cancer, e.g., non-small cell lung cancer.In one embodiment, the anti-TIM-3 antibody molecule is used withplatinum doublet therapy to treat lung cancer.

In yet another embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is used to treat a renal cancer, e.g.,renal cell carcinoma (RCC) (e.g., clear cell renal cell carcinoma(CCRCC) or metastatic RCC. The anti-TIM-3 antibody molecule can beadministered in combination with one or more of: an immune-basedstrategy (e.g., interleukin-2 or interferon-α), a targeted agent (e.g.,a VEGF inhibitor such as a monoclonal antibody to VEGF); a VEGF tyrosinekinase inhibitor such as sunitinib, sorafenib, axitinib and pazopanib;an RNAi inhibitor), or an inhibitor of a downstream mediator of VEGFsignaling, e.g., an inhibitor of the mammalian target of rapamycin(mTOR), e.g., everolimus and temsirolimus.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules described herein, alone or in combinationwith another immunomodulator (e.g., an anti-LAG-3, anti-PD-1 oranti-PD-L1 antibody molecule), for treatment of pancreatic cancerincludes, but is not limited to, a chemotherapeutic agent, e.g.,paclitaxel or a paclitaxel agent (e.g., a paclitaxel formulation such asTAXOL, an albumin-stabilized nanoparticle paclitaxel formulation (e.g.,ABRAXANE) or a liposomal paclitaxel formulation); gemcitabine (e.g.,gemcitabine alone or in combination with AXP107-11); otherchemotherapeutic agents such as oxaliplatin, 5-fluorouracil,capecitabine, rubitecan, epirubicin hydrochloride, NC-6004, cisplatin,docetaxel (e.g., TAXOTERE), mitomycin C, ifosfamide; interferon;tyrosine kinase inhibitor (e.g., EGFR inhibitor (e.g., erlotinib,panitumumab, cetuximab, nimotuzumab); HER2/neu receptor inhibitor (e.g.,trastuzumab); dual kinase inhibitor (e.g., bosutinib, saracatinib,lapatinib, vandetanib); multikinase inhibitor (e.g., sorafenib,sunitinib, XL184, pazopanib); VEGF inhibitor (e.g., bevacizumab, AV-951,brivanib); radioimmunotherapy (e.g., XR303); cancer vaccine (e.g., GVAX,survivin peptide); COX-2 inhibitor (e.g., celecoxib); IGF-1 receptorinhibitor (e.g., AMG 479, MK-0646); mTOR inhibitor (e.g., everolimus,temsirolimus); IL-6 inhibitor (e.g., CNTO 328); cyclin-dependent kinaseinhibitor (e.g., P276-00, UCN-01); Altered Energy Metabolism-Directed(AEMD) compound (e.g., CPI-613); HDAC inhibitor (e.g., vorinostat);TRAIL receptor 2 (TR-2) agonist (e.g., conatumumab); MEK inhibitor(e.g., AS703026, selumetinib, GSK1120212); Raf/MEK dual kinase inhibitor(e.g., RO5126766); Notch signaling inhibitor (e.g., MK0752); monoclonalantibody-antibody fusion protein (e.g., L19IL2); curcumin; HSP90inhibitor (e.g., tanespimycin, STA-9090); rIL-2; denileukin diftitox;topoisomerase 1 inhibitor (e.g., irinotecan, PEP02); statin (e.g.,simvastatin); Factor VIIa inhibitor (e.g., PCI-27483); AKT inhibitor(e.g., RX-0201); hypoxia-activated prodrug (e.g., TH-302); metforminhydrochloride, gamma-secretase inhibitor (e.g., R04929097);ribonucleotide reductase inhibitor (e.g., 3-AP); immunotoxin (e.g.,HuC242-DM4); PARP inhibitor (e.g., KU-0059436, veliparib); CTLA-4inhbitor (e.g., CP-675,206, ipilimumab); AdV-tk therapy; proteasomeinhibitor (e.g., bortezomib (Velcade), NPI-0052); thiazolidinedione(e.g., pioglitazone); NPC-1C; Aurora kinase inhibitor (e.g.,R763/AS703569), CTGF inhibitor (e.g., FG-3019); siG12D LODER; andradiation therapy (e.g., tomotherapy, stereotactic radiation, protontherapy), surgery, and a combination thereof. In certain embodiments, acombination of paclitaxel or a paclitaxel agent, and gemcitabine can beused with the anti-TIM-3 antibody molecules described herein.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibodymolecule), for treatment of small cell lung cancer includes, but is notlimited to, a chemotherapeutic agent, e.g., etoposide, carboplatin,cisplatin, oxaliplatin, irinotecan, topotecan, gemcitabine, liposomalSN-38, bendamustine, temozolomide, belotecan, NK012, FR901228,flavopiridol); tyrosine kinase inhibitor (e.g., EGFR inhibitor (e.g.,erlotinib, gefitinib, cetuximab, panitumumab); multikinase inhibitor(e.g., sorafenib, sunitinib); VEGF inhibitor (e.g., bevacizumab,vandetanib); cancer vaccine (e.g., GVAX); Bcl-2 inhibitor (e.g.,oblimersen sodium, ABT-263); proteasome inhibitor (e.g., bortezomib(Velcade), NPI-0052), paclitaxel or a paclitaxel agent; docetaxel; IGF-1receptor inhibitor (e.g., AMG 479); HGF/SF inhibitor (e.g., AMG 102,MK-0646); chloroquine; Aurora kinase inhibitor (e.g., MLN8237);radioimmunotherapy (e.g., TF2); HSP90 inhibitor (e.g., tanespimycin,STA-9090); mTOR inhibitor (e.g., everolimus); Ep-CAM-/CD3-bispecificantibody (e.g., MT110); CK-2 inhibitor (e.g., CX-4945); HDAC inhibitor(e.g., belinostat); SMO antagonist (e.g., BMS 833923); peptide cancervaccine, and radiation therapy (e.g., intensity-modulated radiationtherapy (IMRT), hypofractionated radiotherapy, hypoxia-guidedradiotherapy), surgery, and combinations thereof. An example of suitabletherapeutics for use in combination with the anti-TIM-3 antibodymolecules, alone or in combination with another immunomodulator (e.g.,an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibody molecule), for treatmentof non-small cell lung cancer includes, but is not limited to, achemotherapeutic agent, e.g., vinorelbine, cisplatin, docetaxel,pemetrexed disodium, etoposide, gemcitabine, carboplatin, liposomalSN-38, TLK286, temozolomide, topotecan, pemetrexed disodium,azacitidine, irinotecan, tegafur-gimeracil-oteracil potassium,sapacitabine); tyrosine kinase inhibitor (e.g., EGFR inhibitor (e.g.,erlotinib, gefitinib, cetuximab, panitumumab, necitumumab, PF-00299804,nimotuzumab, RO5083945), MET inhibitor (e.g., PF-02341066, ARQ 197),PI3K kinase inhibitor (e.g., XL147, GDC-0941), Raf/MEK dual kinaseinhibitor (e.g., RO5126766), PI3K/mTOR dual kinase inhibitor (e.g.,XL765), SRC inhibitor (e.g., dasatinib), dual inhibitor (e.g., BIBW2992, GSK1363089, ZD6474, AZD0530, AG-013736, lapatinib, MEHD7945A,linifanib), multikinase inhibitor (e.g., sorafenib, sunitinib,pazopanib, AMG 706, XL184, MGCD265, BMS-690514, R935788), VEGF inhibitor(e.g., endostar, endostatin, bevacizumab, cediranib, BIBF 1120,axitinib, tivozanib, AZD2171), cancer vaccine (e.g., BLP25 liposomevaccine, GVAX, recombinant DNA and adenovirus expressing L523S protein),Bcl-2 inhibitor (e.g., oblimersen sodium), proteasome inhibitor (e.g.,bortezomib, carfilzomib, NPI-0052, MLN9708), paclitaxel or a paclitaxelagent, docetaxel, IGF-1 receptor inhibitor (e.g., cixutumumab, MK-0646,OSI 906, CP-751,871, BIIB022), hydroxychloroquine, HSP90 inhibitor(e.g., tanespimycin, STA-9090, AUY922, XL888), mTOR inhibitor (e.g.,everolimus, temsirolimus, ridaforolimus), Ep-CAM-/CD3-bispecificantibody (e.g., MT110), CK-2 inhibitor (e.g., CX-4945), HDAC inhibitor(e.g., MS 275, LBH589, vorinostat, valproic acid, FR901228), DHFRinhibitor (e.g., pralatrexate), retinoid (e.g., bexarotene, tretinoin),antibody-drug conjugate (e.g., SGN-15), bisphosphonate (e.g., zoledronicacid), cancer vaccine (e.g., belagenpumatucel-L), low molecular weightheparin (LMWH) (e.g., tinzaparin, enoxaparin), GSK1572932A, melatonin,talactoferrin, dimesna, topoisomerase inhibitor (e.g., amrubicin,etoposide, karenitecin), nelfinavir, cilengitide, ErbB3 inhibitor (e.g.,MM-121, U3-1287), survivin inhibitor (e.g., YM155, LY2181308), eribulinmesylate, COX-2 inhibitor (e.g., celecoxib), pegfilgrastim, Polo-likekinase 1 inhibitor (e.g., BI 6727), TRAIL receptor 2 (TR-2) agonist(e.g., CS-1008), CNGRC peptide (SEQ ID NO: 225)-TNF alpha conjugate,dichloroacetate (DCA), HGF inhibitor (e.g., SCH 900105), SAR240550,PPAR-gamma agonist (e.g., CS-7017), gamma-secretase inhibitor (e.g.,RO4929097), epigenetic therapy (e.g., 5-azacitidine), nitroglycerin, MEKinhibitor (e.g., AZD6244), cyclin-dependent kinase inhibitor (e.g.,UCN-01), cholesterol-Fus1, antitubulin agent (e.g., E7389),farnesyl-OH-transferase inhibitor (e.g., lonafarnib), immunotoxin (e.g.,BB-10901, SS1 (dsFv) PE38), fondaparinux, vascular-disrupting agent(e.g., AVE8062), PD-L1 inhibitor (e.g., MDX-1105, MDX-1106),beta-glucan, NGR-hTNF, EMD 521873, MEK inhibitor (e.g., GSK1120212),epothilone analog (e.g., ixabepilone), kinesin-spindle inhibitor (e.g.,4SC-205), telomere targeting agent (e.g., KML-001), P70 pathwayinhibitor (e.g., LY2584702), AKT inhibitor (e.g., MK-2206), angiogenesisinhibitor (e.g., lenalidomide), Notch signaling inhibitor (e.g.,OMP-21M18), radiation therapy, surgery, and combinations thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibodymolecule), for treatment of ovarian cancer includes, but is not limitedto, a chemotherapeutic agent (e.g., paclitaxel or a paclitaxel agent;docetaxel; carboplatin; gemcitabine; doxorubicin; topotecan; cisplatin;irinotecan, TLK286, ifosfamide, olaparib, oxaliplatin, melphalan,pemetrexed disodium, SJG-136, cyclophosphamide, etoposide, decitabine);ghrelin antagonist (e.g., AEZS-130), immunotherapy (e.g., APC8024,oregovomab, OPT-821), tyrosine kinase inhibitor (e.g., EGFR inhibitor(e.g., erlotinib), dual inhibitor (e.g., E7080), multikinase inhibitor(e.g., AZD0530, JI-101, sorafenib, sunitinib, pazopanib), ON 01910.Na),VEGF inhibitor (e.g., bevacizumab, BIBF 1120, cediranib, AZD2171), PDGFRinhibitor (e.g., IMC-3G3), paclitaxel, topoisomerase inhibitor (e.g.,karenitecin, Irinotecan), HDAC inhibitor (e.g., valproate, vorinostat),folate receptor inhibitor (e.g., farletuzumab), angiopoietin inhibitor(e.g., AMG 386), epothilone analog (e.g., ixabepilone), proteasomeinhibitor (e.g., carfilzomib), IGF-1 receptor inhibitor (e.g., OSI 906,AMG 479), PARP inhibitor (e.g., veliparib, AG014699, iniparib, MK-4827),Aurora kinase inhibitor (e.g., MLN8237, ENMD-2076), angiogenesisinhibitor (e.g., lenalidomide), DHFR inhibitor (e.g., pralatrexate),radioimmunotherapeutic agnet (e.g., Hu3S193), statin (e.g., lovastatin),topoisomerase 1 inhibitor (e.g., NKTR-102), cancer vaccine (e.g., p53synthetic long peptides vaccine, autologous OC-DC vaccine), mTORinhibitor (e.g., temsirolimus, everolimus), BCR/ABL inhibitor (e.g.,imatinib), ET-A receptor antagonist (e.g., ZD4054), TRAIL receptor 2(TR-2) agonist (e.g., CS-1008), HGF/SF inhibitor (e.g., AMG 102),EGEN-001, Polo-like kinase 1 inhibitor (e.g., BI 6727), gamma-secretaseinhibitor (e.g., R04929097), Wee-1 inhibitor (e.g., MK-1775),antitubulin agent (e.g., vinorelbine, E7389), immunotoxin (e.g.,denileukin diftitox), SB-485232, vascular-disrupting agent (e.g.,AVE8062), integrin inhibitor (e.g., EMD 525797), kinesin-spindleinhibitor (e.g., 4SC-205), revlimid, HER2 inhibitor (e.g., MGAH22),ErrB3 inhibitor (e.g., MM-121), radiation therapy; and combinationsthereof.

In one exemplary embodiment, the anti-TIM-3 antibody molecule, alone orin combination with another immunomodulator (e.g., an anti-LAG-3,anti-PD-1 or anti-PD-L1 antibody molecule), is used to treat a myeloma,alone or in combination with one or more of: chemotherapy or otheranti-cancer agents (e.g., thalidomide analogs, e.g., lenalidomide), HSCT(Cook, R. (2008) J Manag Care Pharm. 14(7 Suppl):19-25), an anti-TIM3antibody (Hallett, W H D et al. (2011) J of American Society for Bloodand Marrow Transplantation 17(8):1133-145), tumor antigen-pulseddendritic cells, fusions (e.g., electrofusions) of tumor cells anddendritic cells, or vaccination with immunoglobulin idiotype produced bymalignant plasma cells (reviewed in Yi, Q. (2009) Cancer J.15(6):502-10).

In yet another embodiment, the anti-TIM-3 antibody molecule, alone or incombination with another immunomodulator (e.g., an anti-LAG-3, anti-PD-1or anti-PD-L1 antibody molecule), is used to treat a renal cancer, e.g.,renal cell carcinoma (RCC) or metastatic RCC. The anti-TIM-3 antibodymolecule can be administered in combination with one or more of: animmune-based strategy (e.g., interleukin-2 or interferon-α), a targetedagent (e.g., a VEGF inhibitor such as a monoclonal antibody to VEGF,e.g., bevacizumab (Rini, B. I. et al. (2010) J. Clin. Oncol.28(13):2137-2143)); a VEGF tyrosine kinase inhibitor such as sunitinib,sorafenib, axitinib and pazopanib (reviewed in Pal. S. K. et al. (2014)Clin. Advances in Hematology & Oncology 12(2):90-99)); an RNAiinhibitor), or an inhibitor of a downstream mediator of VEGF signaling,e.g., an inhibitor of the mammalian target of rapamycin (mTOR), e.g.,everolimus and temsirolimus (Hudes, G. et al. (2007) N. Engl. J. Med.356(22):2271-2281, Motzer, R. J. et al. (2008) Lancet 372: 449-456).

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules described herein, alone or in combinationwith another immunomodulator (e.g., an anti-LAG-3, anti-PD-1 oranti-PD-L1 antibody molecule), for treatment of chronic myelogenousleukemia (AML) according to the invention includes, but is not limitedto, a chemotherapeutic (e.g., cytarabine, hydroxyurea, clofarabine,melphalan, thiotepa, fludarabine, busulfan, etoposide, cordycepin,pentostatin, capecitabine, azacitidine, cyclophosphamide, cladribine,topotecan), tyrosine kinase inhibitor (e.g., BCR/ABL inhibitor (e.g.,imatinib, nilotinib), ON 01910.Na, dual inhibitor (e.g., dasatinib,bosutinib), multikinase inhibitor (e.g., DCC-2036, ponatinib, sorafenib,sunitinib, RGB-286638)), interferon alfa, steroids, apoptotic agent(e.g., omacetaxine mepesuccinat), immunotherapy (e.g., allogeneic CD4+memory Th1-like T cells/microparticle-bound anti-CD3/anti-CD28,autologous cytokine induced killer cells (CIK), AHN-12), CD52 targetingagent (e.g., alemtuzumab), HSP90 inhibitor (e.g., tanespimycin,STA-9090, AUY922, XL888), mTOR inhibitor (e.g., everolimus), SMOantagonist (e.g., BMS 833923), ribonucleotide reductase inhibitor (e.g.,3-AP), JAK-2 inhibitor (e.g., INCB018424), Hydroxychloroquine, retinoid(e.g., fenretinide), cyclin-dependent kinase inhibitor (e.g., UCN-01),HDAC inhibitor (e.g., belinostat, vorinostat, JNJ-26481585), PARPinhibitor (e.g., veliparib), MDM2 antagonist (e.g., RO5045337), Aurora Bkinase inhibitor (e.g., TAK-901), radioimmunotherapy (e.g.,actinium-225-labeled anti-CD33 antibody HuM195), Hedgehog inhibitor(e.g., PF-04449913), STAT3 inhibitor (e.g., OPB-31121), KB004, cancervaccine (e.g., AG858), bone marrow transplantation, stem celltransplantation, radiation therapy, and combinations thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibodymolecule), for treatment of chronic lymphocytic leukemia (CLL) includes,but is not limited to, a chemotherapeutic agent (e.g., fludarabine,cyclophosphamide, doxorubicin, vincristine, chlorambucil, bendamustine,chlorambucil, busulfan, gemcitabine, melphalan, pentostatin,mitoxantrone, 5-azacytidine, pemetrexed disodium), tyrosine kinaseinhibitor (e.g., EGFR inhibitor (e.g., erlotinib), BTK inhibitor (e.g.,PCI-32765), multikinase inhibitor (e.g., MGCD265, RGB-286638), CD-20targeting agent (e.g., rituximab, ofatumumab, RO5072759, LFB-R603), CD52targeting agent (e.g., alemtuzumab), prednisolone, darbepoetin alfa,lenalidomide, Bcl-2 inhibitor (e.g., ABT-263), immunotherapy (e.g.,allogeneic CD4+ memory Th1-like T cells/microparticle-boundanti-CD3/anti-CD28, autologous cytokine induced killer cells (CIK)),HDAC inhibitor (e.g., vorinostat, valproic acid, LBH589, JNJ-26481585,AR-42), XIAP inhibitor (e.g., AEG35156), CD-74 targeting agent (e.g.,milatuzumab), mTOR inhibitor (e.g., everolimus), AT-101, immunotoxin(e.g., CAT-8015, anti-Tac(Fv)-PE38 (LMB-2)), CD37 targeting agent (e.g.,TRU-016), radioimmunotherapy (e.g., 131-tositumomab),hydroxychloroquine, perifosine, SRC inhibitor (e.g., dasatinib),thalidomide, PI3K delta inhibitor (e.g., CAL-101), retinoid (e.g.,fenretinide), MDM2 antagonist (e.g., RO5045337), plerixafor, Aurorakinase inhibitor (e.g., MLN8237, TAK-901), proteasome inhibitor (e.g.,bortezomib), CD-19 targeting agent (e.g., MEDI-551, MOR208), MEKinhibitor (e.g., ABT-348), JAK-2 inhibitor (e.g., INCB018424),hypoxia-activated prodrug (e.g., TH-302), paclitaxel or a paclitaxelagent, HSP90 inhibitor, AKT inhibitor (e.g., MK2206), HMG-CoA inhibitor(e.g., simvastatin), GNKG186, radiation therapy, bone marrowtransplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules described herein, alone or in combinationwith another immunomodulator (e.g., an anti-LAG-3, anti-PD-1 oranti-PD-L1 antibody molecule), for treatment of acute lymphocyticleukemia (ALL) includes, but is not limited to, a chemotherapeutic agent(e.g., prednisolone, dexamethasone, vincristine, asparaginase,daunorubicin, cyclophosphamide, cytarabine, etoposide, thioguanine,mercaptopurine, clofarabine, liposomal annamycin, busulfan, etoposide,capecitabine, decitabine, azacitidine, topotecan, temozolomide),tyrosine kinase inhibitor (e.g., BCR/ABL inhibitor (e.g., imatinib,nilotinib), ON 01910.Na, multikinase inhibitor (e.g., sorafenib)), CD-20targeting agent (e.g., rituximab), CD52 targeting agent (e.g.,alemtuzumab), HSP90 inhibitor (e.g., STA-9090), mTOR inhibitor (e.g.,everolimus, rapamycin), JAK-2 inhibitor (e.g., INCB018424), HER2/neureceptor inhibitor (e.g., trastuzumab), proteasome inhibitor (e.g.,bortezomib), methotrexate, asparaginase, CD-22 targeting agent (e.g.,epratuzumab, inotuzumab), immunotherapy (e.g., autologous cytokineinduced killer cells (CIK), AHN-12), blinatumomab, cyclin-dependentkinase inhibitor (e.g., UCN-01), CD45 targeting agent (e.g., BC8), MDM2antagonist (e.g., RO5045337), immunotoxin (e.g., CAT-8015, DT2219ARL),HDAC inhibitor (e.g., JNJ-26481585), JVRS-100, paclitaxel or apaclitaxel agent, STAT3 inhibitor (e.g., OPB-31121), PARP inhibitor(e.g., veliparib), EZN-2285, radiation therapy, steroid, bone marrowtransplantation, stem cell transplantation, or a combination thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules described herein, alone or in combinationwith another immunomodulator (e.g., an anti-LAG-3, anti-PD-1 oranti-PD-L1 antibody molecule), for treatment of acute myeloid leukemia(AML) includes, but is not limited to, a chemotherapeutic agent (e.g.,cytarabine, daunorubicin, idarubicin, clofarabine, decitabine,vosaroxin, azacitidine, clofarabine, ribavirin, CPX-351, treosulfan,elacytarabine, azacitidine), tyrosine kinase inhibitor (e.g., BCR/ABLinhibitor (e.g., imatinib, nilotinib), ON 01910.Na, multikinaseinhibitor (e.g., midostaurin, SU 11248, quizartinib, sorafinib)),immunotoxin (e.g., gemtuzumab ozogamicin), DT3881L3 fusion protein, HDACinhibitor (e.g., vorinostat, LBH589), plerixafor, mTOR inhibitor (e.g.,everolimus), SRC inhibitor (e.g., dasatinib), HSP90 inhbitor (e.g.,STA-9090), retinoid (e.g., bexarotene, Aurora kinase inhibitor (e.g., BI811283), JAK-2 inhibitor (e.g., INCB018424), Polo-like kinase inhibitor(e.g., BI 6727), cenersen, CD45 targeting agent (e.g., BC8),cyclin-dependent kinase inhibitor (e.g., UCN-01), MDM2 antagonist (e.g.,RO5045337), mTOR inhibitor (e.g., everolimus), LY573636-sodium, ZRx-101,MLN4924, lenalidomide, immunotherapy (e.g., AHN-12), histaminedihydrochloride, radiation therapy, bone marrow transplantation, stemcell transplantation, and a combination thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules described herein, alone or in combinationwith another immunomodulator (e.g., an anti-LAG-3, anti-PD-1 oranti-PD-L1 antibody molecule), for treatment of multiple myeloma (MM)includes, but is not limited to, a chemotherapeutic agent (e.g.,melphalan, amifostine, cyclophosphamide, doxorubicin, clofarabine,bendamustine, fludarabine, adriamycin, SyB L-0501), thalidomide,lenalidomide, dexamethasone, prednisone, pomalidomide, proteasomeinhibitor (e.g., bortezomib, carfilzomib, MLN9708), cancer vaccine(e.g., GVAX), CD-40 targeting agent (e.g., SGN-40, CHIR-12.12),perifosine, zoledronic acid, Immunotherapy (e.g., MAGE-A3, NY-ESO-1,HuMax-CD38), HDAC inhibitor (e.g., vorinostat, LBH589, AR-42), aplidin,cycline-dependent kinase inhibitor (e.g., PD-0332991, dinaciclib),arsenic trioxide, CB3304, HSP90 inhibitor (e.g., KW-2478), tyrosinekinase inhibitor (e.g., EGFR inhibitor (e.g., cetuximab), multikinaseinhibitor (e.g., AT9283)), VEGF inhibitor (e.g., bevacizumab),plerixafor, MEK inhibitor (e.g., AZD6244), IPH2101, atorvastatin,immunotoxin (e.g., BB-10901), NPI-0052, radioimmunotherapeutic (e.g.,yttrium Y 90 ibritumomab tiuxetan), STAT3 inhibitor (e.g., OPB-31121),MLN4924, Aurora kinase inhibitor (e.g., ENMD-2076), IMGN901, ACE-041,CK-2 inhibitor (e.g., CX-4945), radiation therapy, bone marrowtransplantation, stem cell transplantation, and a combination thereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-1 or anti-PD-L1 antibodymolecule), for treatment of prostate cancer includes, but is not limitedto, a chemotherapeutic agent (e.g., docetaxel, carboplatin,fludarabine), abiraterone, hormonal therapy (e.g., flutamide,bicalutamide, nilutamide, cyproterone acetate, ketoconazole,aminoglutethimide, abarelix, degarelix, leuprolide, goserelin,triptorelin, buserelin), tyrosine kinase inhibitor (e.g., dual kinaseinhibitor (e.g., lapatanib), multikinase inhibitor (e.g., sorafenib,sunitinib)), VEGF inhibitor (e.g., bevacizumab), TAK-700, cancer vaccine(e.g., BPX-101, PEP223), lenalidomide, TOK-001, IGF-1 receptor inhibitor(e.g., cixutumumab), TRC105, Aurora A kinase inhibitor (e.g., MLN8237),proteasome inhibitor (e.g., bortezomib), OGX-011, radioimmunotherapy(e.g., HuJ591-GS), HDAC inhibitor (e.g., valproic acid, SB939, LBH589),hydroxychloroquine, mTOR inhibitor (e.g., everolimus), dovitiniblactate, diindolylmethane, efavirenz, OGX-427, genistein, IMC-3G3,bafetinib, CP-675,206, radiation therapy, surgery, or a combinationthereof.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of HNSCC includes, but is not limited to, oneor both of Compound A8 as described herein (or a compound described inPCT Publication No. WO2010/029082) and cetuximab (e.g., Erbitux,marketed by BMS). In some embodiments, the therapeutic (e.g., theCompound A8 or compound related to A8) is a PI3K modulator, e.g., a PI3Kinhibitor. In some embodiments, the therapeutic (e.g., cetuximab)modulates, e.g., inhibits, EGFR. In some embodiments, the cancer has, oris identified as having, elevated levels or activity of PI3K or EGFRcompared to a control cell or reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of gastric cancer, e.g., MSI-high and/or EBV+gastric cancer, includes, but is not limited to, Compound A8 asdescribed herein (or a compound described in PCT Publication No.WO2010/029082). In some embodiments, the therapeutic (e.g., the CompoundA8 or compound related to A8) is a PI3K modulator, e.g., a PI3Kinhibitor. In some embodiments, the cancer has, or is identified ashaving, elevated levels or activity of PI3K compared to a control cellor reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of gastric cancer, e.g., MSI-high and/orRNF43-inactivated gastric cancer, includes, but is not limited to,Compound A28 as described herein (or a compound described in PCTPublication No. WO2010/101849). In some embodiments, the therapeutic(e.g., the Compound A28 or compound related to A28) is a modulator,e.g., inhibitor, of porcupine. In some embodiments, the cancer has, oris identified as having, elevated levels or activity of porcupinecompared to a control cell or reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of GI stromal tumor (GIST), includes, but isnot limited to, Compound A16 as described herein (or a compounddescribed in PCT Publication No. WO1999/003854). In some embodiments,the therapeutic (e.g., the Compound A16 or compound related to A16) is amodulator, e.g., inhibitor, of a tyrosine kinase. In some embodiments,the cancer has, or is determined to have, elevated levels or activity ofa tyrosine kinase compared to a control cell or reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of NSCLC, e.g., squamous or adenocarcinoma,includes, but is not limited to, one or both of Compound A17 asdescribed herein (or a compound described in U.S. Pat. Nos. 7,767,675and 8,420,645) and Compound A23 as described herein (or a compounddescribed in PCT Publication No. WO2003/077914). In some embodiments,the compound (e.g., the Compound A17 or compound related to A17)modulates, e.g., inhibits, c-MET. In some embodiments, the compound(e.g., the Compound A23 or compound related to A23) modulates, e.g.,inhibits, Alk. In some embodiments, the cancer has, or is determined tohave, elevated levels or activity of one or both of c-MET or Alkcompared to a control cell or reference value. In some embodiments, thecancer has, or is identified as having, a mutation in EGFR.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of melanoma (e.g., NRAS melanoma) includes, butis not limited to, one or both of Compound A24 as described herein (or acompound described in U.S. Pat. Nos. 8,415,355 and 8,685,980) andCompound A34 as described herein (or a compound described in PCTPublication No. WO2003/077914). In some embodiments, the compound (e.g.,the Compound A24 or compound related to A24) modulates, e.g., inhibits,one or more of JAK and CDK4/6. In some embodiments, the compound (e.g.,the Compound A34 or compound related to A34) modulates, e.g., inhibits,MEK. In some embodiments, the cancer has, or is identified as having,elevated levels or activity of one or more of JAK, CDK4/6, and MEKcompared to a control cell or reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of melanoma (e.g., NRAS melanoma) includes, butis not limited to, one or both of Compound A29 as described herein (or acompound described in PCT Publication No. WO2011/025927) and CompoundA34 as described herein (or a compound described in PCT Publication No.WO2003/077914). In some embodiments, the compound (e.g., the CompoundA29 or compound related to A29) modulates, e.g., inhibits, BRAF. In someembodiments, the compound (e.g., the Compound A34 or compound related toA34) modulates, e.g., inhibits, MEK. In some embodiments, the cancerhas, or is identified as having, elevated levels or activity of one orboth of BRAF and MEK compared to a control cell or reference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), for treatment of squamous NSCLC includes, but is not limitedto, Compound A5 as described herein (or a compound described in U.S.Pat. No. 8,552,002). In some embodiments, the compound (e.g., theCompound A5 or compound related to A5) modulates, e.g., inhibits, FGFR.In some embodiments, the cancer has, or is identified as having,elevated levels or activity of FGFR compared to a control cell orreference value.

An example of suitable therapeutics for use in combination with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-Ll or anti-PD-1 antibodymolecule), for treatment of colorectal cancer includes, but is notlimited to, one or both of Compound A29 as described herein (or acompound PCT Publication No. WO2011/025927) and cetuximab (e.g.,Erbitux, marketed by BMS). In some embodiments, the therapeutic (e.g.,the Compound A29 or compound related to A29) modulates, e.g., inhibits,BRAF. In some embodiments, the therapeutic (e.g., cetuximab) modulates,e.g., inhibits EGFR. In some embodiments, the cancer has, or isidentified as having, elevated levels or activity of BRAF or EGFRcompared to a control cell or reference value.

This disclosure also provides a method of treating cancer with CompoundA8, cetuximab, and a TIM-3 antibody molecule (optionally in combinationwith a PD-1 antibody molecule or LAG-3 antibody molecule). In someembodiments, the patient is first treated with Compound A8 andcetuximab. This treatment continues for an amount of time, e.g., apredetermined amount of time, e.g., about 1, 2, 4, 6, 8, 10, or 12months. Next, the TIM-3 antibody molecule (optionally in combinationwith a PD-1 antibody molecule or LAG-3 antibody molecule) isadministered. The TIM-3 antibody can optionally be administered incombination with cetuximab.

In some embodiments, the patient is first treated with all three ofCompound A8, cetuximab, and a TIM-3 antibody molecule (optionally incombination with a PD-1 antibody molecule or LAG-3 antibody molecule).This treatment continues for an amount of time, e.g., a predeterminedamount of time, e.g., about 6, 8, 10, or 12 months. Next, the CompoundA8 and/or cetuximab can be tapered off, so that the maintenance phaseinvolves treatment with the TIM-3 antibody molecule (e.g., as amonotherapy, or in combination with a PD-1 antibody molecule or LAG-3antibody molecule) but not Compound A8 or cetuximab.

In other embodiments, the three compounds (Compound A8, cetuximab, and aTIM-3 antibody molecule, optionally in combination with a PD-1 antibodymolecule or LAG-3 antibody molecule) are given sequentially at theoutset of the treatment. For instance, Compound A8 and cetuximab can begiven first, as described above. Next, the TIM-3 antibody molecule(optionally in combination with a PD-1 antibody molecule or LAG-3antibody molecule) is added to the regimen. Next, the Compound A8 and/orcetuximab can be tapered off as described above.

Exemplary doses for the three (or more) agent regimens are as follows.The TIM-3 antibody molecule can be administered, e.g., at a dose ofabout 1 to 40 mg/kg, e.g., 1 to 30 mg/kg, e.g., about 5 to 25 mg/kg,about 10 to 20 mg/kg, about 1 to 5 mg/kg, or about 3 mg/kg. In someembodiments, the Compound A8 is administered at a dose of approximately200-300, 300-400, or 200-400 mg. In some embodiments, the cetuximab isadministered at a 400 mg/m2 initial dose as a 120-minute intravenousinfusion followed by 250 mg/m2 weekly infused over 60 minutes. Inembodiments, one or more of the Compound A8, cetuximab, and TIM-3antibody molecule is administered at a dose that is lower than the doseat which that agent is typically administered as a monotherapy, e.g.,about 0-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or80-90% lower than the dose at which that agent is typically administeredas a monotherapy. In embodiments, the one or more of the Compound A8,cetuximab, and TIM-3 antibody molecule is administered at a dose that islower than the dose of that agent recited in this paragraph, e.g., about0-10%, 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90%lower than the dose of that agent recited in this paragraph. In certainembodiments, the concentration of the Compound A8 that is required toachieve inhibition, e.g., growth inhibition, is lower when the CompoundA8 is administered in combination with one or both of the cetuximab andTIM-3 antibody molecule than when the Compound A8 is administeredindividually. In certain embodiments, the concentration of the cetuximabthat is required to achieve inhibition, e.g., growth inhibition, islower when the cetuximab is administered in combination with one or bothof the Compound A8 and TIM-3 antibody molecule than when the cetuximabis administered individually. In certain embodiments, the concentrationof the TIM-3 antibody molecule that is required to achieve inhibition,e.g., growth inhibition, is lower when the TIM-3 antibody molecule isadministered in combination with one or both of the cetuximab andCompound A8 than when the TIM-3 antibody molecule is administeredindividually.

Additionally disclosed herein is a method of treating cancer with theanti-TIM-3 antibody molecules, alone or in combination with anotherimmunomodulator (e.g., an anti-LAG-3, anti-PD-L1 or anti-PD-1 antibodymolecule), and a targeted anti-cancer agent, e.g., an agent that targetsone or more proteins. In some embodiments, the anti-TIM-3 antibodymolecule (and optionally other immunomodulator(s)) are administeredfirst, and the targeted anti-cancer agent is administered second. Thelength of time between administration of the anti-TIM-3 antibodymolecule and the targeted anti-cancer agent can be, e.g., 10, 20, or 30minutes, 1, 2, 4, 6, or 12 hours, or 1, 2, 3, 4, 5, 6, or 7 days, or anyspan of time within this range. In certain embodiments, the anti-TIM-3antibody molecule is administered repeatedly over a period of time(e.g., 1, 2, 3, 4, 5, or 6 days, or 1, 2, 4, 8, 12, 16, or 20 weeks, orany span of time within this range) before the targeted anti-canceragent is administered. In other embodiments, the anti-TIM-3 antibodymolecule and the targeted anti-cancer agent are administered atsubstantially the same time.

Methods of Treating Infectious Diseases

Other methods of the invention are used to treat patients that have beenexposed to particular toxins or pathogens. Based on, at least, theExamples herein, anti-TIM-3 antibodies can stimulate NK cell mediatedkilling of target cells and can enhances IFN-gamma secretion andproliferation of CD4+ T cells. Accordingly, in certain embodiments, theanti-TIM-3 antibody molecules described herein are suitable for use instimulating an immune response against an infectious agent. Accordingly,another aspect of the invention provides a method of treating aninfectious disease in a subject comprising administering to the subjectan anti-TIM-3 antibody molecule, such that the subject is treated forthe infectious disease. In the treatment of infection (e.g., acuteand/or chronic), administration of the anti-TIM-3 antibody molecules canbe combined with conventional treatments in addition to or in lieu ofstimulating natural host immune defenses to infection. Natural hostimmune defenses to infection include, but are not limited toinflammation, fever, antibody-mediated host defense,T-lymphocyte-mediated host defenses, including lymphokine secretion andcytotoxic T-cells (especially during viral infection), complementmediated lysis and opsonization (facilitated phagocytosis), andphagocytosis. The ability of the anti-TIM-3 antibody molecules toreactivate dysfunctional T-cells would be useful to treat chronicinfections, in particular those in which cell-mediated immunity isimportant for complete recovery.

Certain methods described herein are used to treat patients that havebeen exposed to particular toxins or pathogens. Some aspects provides amethod of treating an infectious disease in a subject comprisingadministering to the subject an anti-TIM-3 antibody molecule, such thatthe subject is treated for the infectious disease.

Similar to its application to tumors as discussed in the previoussection, aIn embodiments, the anti-TIM-3 antibody molecules can be usedalone, or as an adjuvant, in combination with vaccines, to stimulate theimmune response to, e.g., pathogens or toxins. Examples of pathogens forwhich this therapeutic approach may be particularly useful, includepathogens for which there is currently no effective vaccine, orpathogens for which conventional vaccines are less than completelyeffective. These include, but are not limited to HIV, Hepatitis (A, B, &C), Influenza, Herpes, Giardia, Malaria, Leishmania, Staphylococcusaureus, Pseudomonas Aeruginosa. Anti-TIM-3 antibody molecule therapy isalso useful against established infections by agents such as HIV thatpresent altered antigens over the course of the infections.

Accordingly, in some embodiments an anti-TIM-3 antibody molecule is usedto treat a subject that has an infection or is at risk of having aninfection. An infection refers to, e.g., a disease or conditionattributable to the presence in a host of a foreign organism or agentthat reproduces within the host. Infections typically involve breach ofa normal mucosal or other tissue barrier by an infectious organism oragent. A subject that has an infection is a subject having objectivelymeasurable infectious organisms or agents present in the subject's body.A subject at risk of having an infection is a subject that ispredisposed to develop an infection. Such a subject can include, forexample, a subject with a known or suspected exposure to an infectiousorganism or agent. A subject at risk of having an infection also caninclude a subject with a condition associated with impaired ability tomount an immune response to an infectious organism or agent, e.g., asubject with a congenital or acquired immunodeficiency, a subjectundergoing radiation therapy or chemotherapy, a subject with a burninjury, a subject with a traumatic injury, a subject undergoing surgeryor other invasive medical or dental procedure.

Infections are broadly classified as bacterial, viral, fungal, orparasitic based on the category of infectious organism or agentinvolved. Other less common types of infection include, e.g., infectionsinvolving rickettsiae, mycoplasmas, and agents causing scrapie, bovinespongiform encephalopthy (BSE), and prion diseases (e.g., kuru andCreutzfeldt-Jacob disease). Examples of bacteria, viruses, fungi, andparasites which cause infection are well known in the art. An infectioncan be acute, subacute, chronic, or latent, and it can be localized orsystemic. Furthermore, an infection can be predominantly intracellularor extracellular during at least one phase of the infectious organism'sor agent's life cycle in the host.

Viruses

Examples of viruses that have been found to cause infections in humansinclude but are not limited to: Retroviridae (e.g., humanimmunodeficiency viruses, such as HIV-1 (also referred to as HTLV-III),HIV-2, LAV or HTLV-III/LAV, or HIV-III, and other isolates, such asHIV-LP; Picornaviridae (e.g., polio viruses, hepatitis A virus;enteroviruses, human Coxsackie viruses, rhinoviruses, echoviruses);Calciviridae (e.g., strains that cause gastroenteritis); Togaviridae(e.g., equine encephalitis viruses, rubella viruses); Flaviviridae(e.g., dengue viruses, encephalitis viruses, yellow fever viruses);Coronaviridae (e.g., coronaviruses); Rhabdoviridae (e.g., vesicularstomatitis viruses, rabies viruses); Filoviridae (e.g., ebola viruses);Paramyxoviridae (e.g., parainfluenza viruses, mumps virus, measlesvirus, respiratory syncytial virus); Orthomyxoviridae (e.g., influenzaviruses); Bungaviridae (e.g., Hantaan viruses, bunga viruses,phleboviruses and Nairo viruses); Arena viridae (hemorrhagic feverviruses); Reoviridae (e.g., reoviruses, orbiviurses and rotaviruses);Birnaviridae; Hepadnaviridae (Hepatitis B virus); Parvoviridae(parvoviruses); Papovaviridae (papilloma viruses, polyoma viruses);Adenoviridae (most adenoviruses); Herpesviridae (herpes simplex virus(HSV) 1 and 2, varicella zoster virus, cytomegalovirus (CMV), herpesvirus; Poxyiridae (variola viruses, vaccinia viruses, pox viruses); andIridoviridae (e.g., African swine fever virus); and unclassified viruses(e.g., the etiological agents of Spongiform encephalopathies, the agentof delta hepatitis (thought to be a defective satellite of hepatitis Bvirus), the agents of non-A, non-B hepatitis (class 1=enterallytransmitted; class 2=parenterally transmitted (i.e., Hepatitis C);Norwalk and related viruses, and astroviruses). Some examples ofpathogenic viruses causing infections treatable by methods hereininclude HIV, hepatitis (A, B, or C), herpes virus (e.g., VZV, HSV-1,HAV-6, HSV-II, and CMV, Epstein Barr virus), adenovirus, influenzavirus, flaviviruses, echovirus, rhinovirus, coxsackie virus, cornovirus,respiratory syncytial virus, mumps virus, rotavirus, measles virus,rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus,papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus andarboviral encephalitis virus.

For infections resulting from viral causes, the anti-TIM-3 antibodymolecules can be combined by application simultaneous with, prior to orsubsequent to application of standard therapies for treating viralinfections. Such standard therapies vary depending upon type of virus,although in almost all cases, administration of human serum containingantibodies (e.g., IgA, IgG) specific to the virus can be effective.

Some examples of pathogenic viruses causing infections treatable bymethods include HIV, hepatitis (A, B, or C), herpes virus (e.g., VZV,HSV-1, HAV-6, HSV-II, and CMV, Epstein Barr virus), adenovirus,influenza virus, flaviviruses, echovirus, rhinovirus, coxsackie virus,cornovirus, respiratory syncytial virus, mumps virus, rotavirus, measlesvirus, rubella virus, parvovirus, vaccinia virus, HTLV virus, denguevirus, papillomavirus, molluscum virus, poliovirus, rabies virus, JCvirus, arboviral encephalitis virus, and ebolaviruses (e.g., BDBV, EBOV,RESTV, SUDV and TAFV).

In one embodiment, the infection is an influenza infection. Influenzainfection can result in fever, cough, myalgia, headache and malaise,which often occur in seasonal epidemics. Influenza is also associatedwith a number of postinfectious disorders, such as encephalitis,myopericarditis, Goodpasture's syndrome, and Reye's syndrome. Influenzainfection also suppresses normal pulmonary antibacterial defenses, suchthat patient's recovering from influenza have an increased risk ofdeveloping bacterial pneumonia. Influenza viral surface proteins showmarked antigenic variation, resulting from mutation and recombination.Thus, cytolytic T lymphocytes are the host's primary vehicle for theelimination of virus after infection. Influenza is classified into threeprimary types: A, B and C. Influenza A is unique in that it infects bothhumans and many other animals (e.g., pigs, horses, birds and seals) andis the principal cause of pandemic influenza. Also, when a cell isinfected by two different influenza A strains, the segmented RNA genomesof two parental virus types mix during replication to create a hybridreplicant, resulting in new epidemic strains. Influenza B does notreplicate in animals and thus has less genetic variation and influenza Chas only a single serotype.

Most conventional therapies are palliatives of the symptoms resultingfrom infection, while the host's immune response actually clears thedisease. However, certain strains (e.g., influenza A) can cause moreserious illness and death. Influenza A may be treated both clinicallyand prophylactically by the administration of the cyclic aminesinhibitors amantadine and rimantadine, which inhibit viral replication.However, the clinical utility of these drugs is limited due to therelatively high incidence of adverse reactions, their narrow anti-viralspectrum (influenza A only), and the propensity of the virus to becomeresistant. The administration of serum IgG antibody to the majorinfluenza surface proteins, hemagglutinin and neuraminidase can preventpulmonary infection, whereas mucosal IgA is required to preventinfection of the upper respiratory tract and trachea. The most effectivecurrent treatment for influenza is vaccination with the administrationof virus inactivated with formalin or β-propiolactone.

In another embodiment, the infection is a hepatitis infection, e.g., aHepatitis B or C infection.

Hepatitis B virus (HB-V) is the most infectious known bloodbornepathogen. It is a major cause of acute and chronic heptatis and hepaticcarcinoma, as well as life-long, chronic infection. Following infection,the virus replicates in hepatocytes, which also then shed the surfaceantigen HBsAg. The detection of excessive levels of HBsAg in serum isused a standard method for diagnosing a hepatitis B infection. An acuteinfection may resolve or it can develop into a chronic persistentinfection. Current treatments for chronic HBV include α-interferon,which increases the expression of class I human leukocyte antigen (HLA)on the surface of hepatocytes, thereby facilitating their recognition bycytotoxic T lymphocytes. Additionally, the nucleoside analogsganciclovir, famciclovir and lamivudine have also shown some efficacy inthe treatment of HBV infection in clinical trials. Additional treatmentsfor HBV include pegylated a-interferon, adenfovir, entecavir andtelbivudine. While passive immunity can be conferred through parentaladministration of anti-HBsAg serum antibodies, vaccination withinactivated or recombinant HBsAg also confers resistance to infection.The anti-TIM-3 antibody molecules may be combined with conventionaltreatments for hepatitis B infections for therapeutic advantage.

Hepatitis C virus (HC-V) infection may lead to a chronic form ofhepatitis, resulting in cirrosis. While symptoms are similar toinfections resulting from Hepatitis B, in distinct contrast to HB-V,infected hosts can be asymptomatic for 10-20 years. The anti-TIM-3antibody molecule can be administered as a monotherapy, or combined withthe standard of care for hepatitis C infection. For example, theanti-TIM-3 antibody molecule can be administered with one or more ofSovaldi (sofosbuvir) Olysio (simeprevir), plus ribavirin or pegylatedinterferon. Although regimens that include Incivek (telaprevir) orVictrelis (boceprevir) plus ribavirin and pegylated interferon are alsoapproved, they are associated with increased side effects and longerduration of treatment and are therefore not considered preferredregimens.

Conventional treatment for HC-V infection includes the administration ofa combination of α-interferon and ribavirin. A promising potentialtherapy for HC-V infection is the protease inhibitor telaprevir(VX-960). Additional treatments include: anti-PD-1 antibody (MDX-1106,Medarex), bavituximab (an antibody that binds anionic phospholipidphosphatidylserine in a B2-glycoprotein I dependent manner, PeregrinePharmaceuticals), anti-HPV viral coat protein E2 antibod(y)(ies) (e.g.,ATL 6865-Ab68+Ab65, XTL Pharmaceuticals) and Civacir® (polyclonalanti-HCV human immune globulin). The anti-PD-L1 antibodies of theinvention may be combined with one or more of these treatments forhepatitis C infections for therapeutic advantage. Protease, polymeraseand NSSA inhibitors which may be used in combination with the anti-TIM-3antibody molecules to specifically treat Hepatitis C infection includethose described in US 2013/0045202, incorporated herein by reference.

In another embodiment, the infection is a measles virus. After anincubation of 9-11 days, hosts infected with the measles virus developfever, cough, coryza and conjunctivitis. Within 1-2 days, anerythematous, maculopapular rash develop, which quickly spreads over theentire body. Because infection also suppresses cellular immunity, thehost is at greater risk for developing bacterial superinfections,including otitis media, pneumonia and postinfectious encephalomyelitis.Acute infection is associated with significant morbidity and mortality,especially in malnourished adolescents.

Treatment for measles includes the passive administration of pooledhuman IgG, which can prevent infection in non-immune subjects, even ifgiven up to one week after exposure. However, prior immunization withlive, attenuated virus is the most effective treatment and preventsdisease in more than 95% of those immunized. As there is one serotype ofthis virus, a single immunization or infection typically results inprotection for life from subsequent infection.

In a small proportion of infected hosts, measles can develop into SSPE,which is a chronic progressive neurologic disorder resulting from apersistent infection of the central nervous system. SSPE is caused byclonal variants of measles virus with defects that interfere with virionassembly and budding. For these patients, reactivation of T-cells withthe anti-TIM-3 antibody molecules so as to facilitate viral clearancewould be desirable.

In another embodiment, the infection is HIV. HIV attacks CD4⁺ cells,including T-lymphocytes, monocyte-macrophages, follicular dendriticcells and Langerhan's cells, and CD4⁺ helper/inducer cells are depleted.As a result, the host acquires a severe defect in cell-mediatedimmunity. Infection with HIV results in AIDS in at least 50% ofindividuals, and is transmitted via sexual contact, administration ofinfected blood or blood products, artificial insemination with infectedsemen, exposure to blood-containing needles or syringes and transmissionfrom an infected mother to infant during childbirth.

A host infected with HIV may be asymptomatic, or may develop an acuteillness that resembling mononucleosis—fever, headache, sore throat,malaise and rash. Symptoms can progress to progressive immunedysfunction, including persistent fever, night sweats, weight loss,unexplained diarrhea, eczema, psoriasis, seborrheic dermatitis, herpeszoster, oral candidiasis and oral hairy leukoplakia. Opportunisticinfections by a host of parasites are common in patients whoseinfections develop into AIDS.

Treatments for HIV include antiviral therapies including nucleosideanalogs, zidovudine (AST) either alone or in combination with didanosineor zalcitabine, dideoxyinosine, dideoxycytidine, lamidvudine, stavudine;reverse transcriptive inhibitors such as delavirdine, nevirapine,loviride, and proteinase inhibitors such as saquinavir, ritonavir,indinavir and nelfinavir. The anti-TIM-3 antibody molecules may becombined with conventional treatments for HIV infections for therapeuticadvantage.

In another embodiment, the infection is a Cytomegalovirus (CMV). CMVinfection is often associated with persistent, latent and recurrentinfection. CMV infects and remains latent in monocytes andgranulocyte-monocyte progenitor cells. The clinical symptoms of CMVinclude mononucleosis-like symptoms (i.e., fever, swollen glands,malaise), and a tendancy to develop allergic skin rashes to antibiotics.The virus is spread by direct contact. The virus is shed in the urine,saliva, semen and to a lesser extent in other body fluids. Transmissioncan also occur from an infected mother to her fetus or newborn and byblood transfusion and organ transplants. CMV infection results ingeneral impairment of cellular immunity, characterized by impairedblastogenic responses to nonspecific mitogens and specific CMV antigens,diminished cytotoxic ability and elevation of CD8 lymphocyte number ofCD4⁺ lymphocytes.

Treatments of CMV infection include the anti-virals ganciclovir,foscarnet and cidovir, but these druges are typically only prescribed inimmunocompromised patients. The anti-TIM-3 antibody molecules may becombined with conventional treatments for cytomegalovirus infections fortherapeutic advantage.

In another embodiment, the infection is Epstein-Barr virus (EBV). EBVcan establish persistent and latent infections and primarily attacks Bcells. Infection with EBV results in the clinical condition ofinfectious mononucleosis, which includes fever, sore throat, often withexudate, generalized lymphadenopathy and splenomegaly. Hepatitis is alsopresent, which can develop into jaundice.

While typical treatments for EBV infections are palliative of symptoms,EBV is associated with the development of certain cancers such asBurkitt's lymphoma and nasopharyngeal cancer. Thus, clearance of viralinfection before these complications result would be of great benefit.The anti-TIM-3 antibody molecules may be combined with conventionaltreatments for Epstein-Barr virus infections for therapeutic advantage.

In another embodiment, the infection is Herpes simplex virus (HSV). HSVis transmitted by direct contact with an infected host. A directinfection may be asymptomatic, but typically result in blisterscontaining infectious particles. The disease manifests as cycles ofactive periods of disease, in which lesions appear and disappear as theviral latently infect the nerve ganglion for subsequent outbreaks.Lesions may be on the face, genitals, eyes and/or hands. In some case,an infection can also cause encephalitis.

Treatments for herpes infections are directed primarily to resolving thesymptomatic outbreaks, and include systemic antiviral medicines such as:acyclovir (e.g., Zovirax®), valaciclovir, famciclovir, penciclovir, andtopical medications such as docosanol (Abreva®), tromantadine andzilactin. The clearance of latent infections of herpes would be of greatclinical benefit. The anti-TIM-3 antibody molecules may be combined withconventional treatments for herpes virus infections for therapeuticadvantage.

In another embodiment, the infection is Human T-lymphotrophic virus(HTLV-1, HTLV-2). HTLV is transmitted via sexual contact, breast feedingor exposure to contaminated blood. The virus activates a subset of T_(H)cells called Th1 cells, resulting in their overproliferation andoverproduction of Th1 related cytokines (e.g., IFN-γ and TNF-α). This inturn results in a suppression of Th2 lymphocytes and reduction of Th2cytokine production (e.g., IL-4, IL-5, IL-10 and IL-13), causing areduction in the ability of an infected host to mount an adequate immuneresponse to invading organisms requiring a Th2-dependent response forclearnance (e.g., parasitic infections, production of mucosal andhumoral antibodies).

HTLV infections cause lead to opportunistic infections resulting inbronchiectasis, dermatitis and superinfections with Staphylococcus spp.and Strongyloides spp. resulting in death from polymicrobial sepsis.HTLV infection can also lead directly to adult T-cell leukemia/lymphomaand progressive demyelinating upper motor neuron disease known asHAM/TSP. The clearance of HTLV latent infections would be of greatclinical benefit. The anti-TIM-3 antibody molecules may be combined withconventional treatments for HTLV infections for therapeutic advantage.

In another embodiment, the infection is Human papilloma virus (HPV). HPVprimarily affects keratinocytes and occurs in two forms: cutaneous andgenital. Transmission is believed to occur through direct contact and/orsexual activity. Both cutaneous and genital HPV infection, can result inwarts and latent infections and sometimes recurring infections, whichare controlled by host immunity which controls the symptoms and blocksthe appearance of warts, but leaves the host capable of transmitting theinfection to others.

Infection with HPV can also lead to certain cancers, such as cervical,anal, vulvar, penile and oropharynial cancer. There are no known curesfor HPV infection, but current treatment is topical application ofImiquimod, which stimulates the immune system to attack the affectedarea. The clearance of HPV latent infections would be of great clinicalbenefit. The anti-TIM-3 antibodies of the invention may be combined withconventional treatments for HPV infections for therapeutic advantage.

In another embodiment, the infection is Ebola virus (EBOV). EBOV is oneof five known viruses within the Ebolavirus genus. EBOV causes severeand often fatal hemorrhagic fever in humans and mammals, known as Ebolavirus disease (EVD). Transmission occurs through contact with blood,secretions, organs, or other boldily fluids of infected patients.Currently, there is no proven treatment or vaccine.

Bacterial Infections

Bacteria include both Gram negative and Gram positive bacteria. Examplesof Gram positive bacteria include, but are not limited to Pasteurellaspecies, Staphylococci species, and Streptococcus species. Examples ofGram negative bacteria include, but are not limited to, Escherichiacoli, Pseudomonas species, and Salmonella species. Specific examples ofinfectious bacteria include but are not limited to: Helicobacterpyloris, Borrelia burgdorferi, Legionella pneumophilia, Mycobacteriaspp. (e.g., M. tuberculosis, M. avium, M. intracellulare, M. kansasii,M. gordonae), Staphylococcus aureus, Neisseria gonorrhoeae, Neisseriameningitidis, Listeria monocytogenes, Streptococcus pyogenes (Group AStreptococcus), Streptococcus agalactiae (Group B Streptococcus),Streptococcus (viridans group), Streptococcus faecalis, Streptococcusbovis, Streptococcus (anaerobic spp.), Streptococcus pneumoniae,pathogenic Campylobacter spp., Enterococcus spp., Haemophilusinfluenzae, Bacillus anthracis, Corynebacterium diphtheriae,Corynebacterium spp., Erysipelothrix rhusiopathiae, Clostridiumperfringens, Clostridium tetani, Enterobacter aerogenes, Klebsiellapneumoniae, Pasteurella multocida, Bacteroides spp., Fusobacteriumnucleatum, Streptobacillus moniliformis, Treponema pallidum, Treponemapertenue, Leptospira, Mycobacterium leprae, Rickettsia, and Actinomycesisraelii. Some examples of pathogenic bacteria causing infectionstreatable by methods herein include chlamydia, rickettsial bacteria,mycobacteria, staphylococci, streptococci, pneumonococci, meningococciand conococci, klebsiella, proteus, serratia, pseudomonas, legionella,diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax,plague, leptospirosis, and Lymes disease bacteria.

Some examples of pathogenic bacteria causing infections treatable bymethods of the invention include syphilis, chlamydia, rickettsialbacteria, mycobacteria, staphylococci, streptococci, pneumonococci,meningococci and conococci, klebsiella, proteus, serratia, pseudomonas,legionella, diphtheria, salmonella, bacilli, cholera, tetanus, botulism,anthrax, plague, leptospirosis, and Lymes disease bacteria. Theanti-TIM-3 antibody molecules can be used in combination with existingtreatment modalities for the aforesaid infections. For example,Treatments for syphilis include penicillin (e.g., penicillin G.),tetracycline, doxycycline, ceftriaxone and azithromycin.

Lyme disease, caused by Borrelia burgdorferi is transmitted into humansthrough tick bites. The disease manifests initially as a localized rash,followed by flu-like symptoms including malaise, fever, headache, stiffneck and arthralgias. Later manifestations can include migratory andpolyarticular arthritis, neurologic and cardiac involvement with cranialnerve palsies and radiculopathy, myocarditis and arrhythmias. Some casesof Lyme disease become persistent, resulting in irreversible damageanalogous to tertiary syphilis. Current therapy for Lyme diseaseincludes primarily the administration of antibiotics.Antibiotic-resistant strains may be treated with hydroxychloroquine ormethotrexate. Antibiotic refractory patients with neuropathic pain canbe treated with gabapentin. Minocycline may be helpful in late/chronicLyme disease with neurological or other inflammatory manifestations.

Other forms of borreliois, such as those resulting from B. recurentis,B. hermsii, B. turicatae, B. parikeri., B. hispanica, B. duttonii and B.persica, as well leptospirosis (E.g., L. interrogans), typically resolvespontaneously unless blood titers reach concentrations to causeintrahepatic obstruction.

Fungi and Parasites

Examples of fungi include: Aspergillus spp., Blastomyces dermatitidis,Candida albicans, other Candida spp., Coccidioides immitis, Cryptococcusneoformans, Histoplasma capsulatum, Chlamydia trachomatis, Nocardiaspp., Pneumocystis carinii. Some examples of pathogenic fungi causinginfections treatable by methods herein include Candida (albicans,krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans,Aspergillus (fumigatus, niger, etc.), Genus Mucorales (mucor, absidia,rhizophus), Sporothrix schenkii, Blastomyces dermatitidis,Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasmacapsulatum.

Parasites include but are not limited to blood-borne and/or tissuesparasites such as Babesia microti, Babesia divergens, Entamoebahistolytica, Giardia lamblia, Leishmania tropica, Leishmania spp.,Leishmania braziliensis, Leishmania donovani, Plasmodium falciparum,Plasmodium malariae, Plasmodium ovale, Plasmodium vivax, and Toxoplasmagondii, Trypanosoma gambiense and Trypanosoma rhodesiense (Africansleeping sickness), Trypanosoma cruzi (Chagas' disease), and Toxoplasmagondii, flat worms, round worms. Some examples of pathogenic parasitescausing infections treatable by methods herein include Entamoebahistolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp.,Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodiumvivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi,Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

Some examples of pathogenic fungi causing infections treatable bymethods of the invention include Candida (albicans, krusei, glabrata,tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus,niger, etc.), Genus Mucorales (mucor, absidia, rhizophus), Sporothrixschenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis,Coccidioides immitis and Histoplasma capsulatum.

Some examples of pathogenic parasites causing infections treatable bymethods described herein include Entamoeba histolytica, Balantidiumcoli, Naegleriafowleri, Acanthamoeba sp., Giardia lambia,Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesiamicroti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani,Toxoplasma gondi, and Nippostrongylus brasiliensis.

In some embodiments, the infectious disease is chosen from hepatitis(e.g., hepatis C infection), or sepsis.

In all of the above methods, anti-TIM-3 antibody molecule therapy can becombined with other forms of immunotherapy such as cytokine treatment(e.g., interferons, GM-CSF, G-CSF, IL-2, IL-21), or bispecific antibodytherapy, which provides for enhanced presentation of tumor antigens(see, e.g., Holliger (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448;Poljak (1994) Structure 2:1121-1123).

Methods of administering various antibody molecules are known in the artand are described below. Suitable dosages of the antibody molecules usedwill depend on the age and weight of the subject and the particular drugused. The antibody molecules can be used as competitive agents forligand binding to inhibit or reduce an undesirable interaction.

The antibody molecules can be used by themselves or conjugated to asecond agent, e.g., a cytotoxic drug, radioisotope, or a protein, e.g.,a protein toxin or a viral protein. This method includes: administeringthe antibody molecule, alone or conjugated to a cytotoxic drug, to asubject requiring such treatment. The antibody molecules can be used todeliver a variety of therapeutic agents, e.g., a cytotoxic moiety, e.g.,a therapeutic drug, a radioisotope, molecules of plant, fungal, orbacterial origin, or biological proteins (e.g., protein toxins) orparticles (e.g., a recombinant viral particles, e.g.; via a viral coatprotein), or mixtures thereof.

Additional Combination Therapies

The anti-TIM-3 antibody molecules can be used in combination with othertherapies. For example, the combination therapy can include ananti-TIM-3 antibody molecule co-formulated with, and/or co-administeredwith, one or more additional therapeutic agents, e.g., one or moreanti-cancer agents, cytotoxic or cytostatic agents, hormone treatment,vaccines, and/or other immunotherapies. In other embodiments, theantibody molecules are administered in combination with othertherapeutic treatment modalities, including surgery, radiation,cryosurgery, and/or thermotherapy. Such combination therapies mayadvantageously utilize lower dosages of the administered therapeuticagents, thus avoiding possible toxicities or complications associatedwith the various monotherapies.

Administered “in combination”, as used herein, means that two (or more)different treatments are delivered to the subject during the course ofthe subject's affliction with the disorder, e.g., the two or moretreatments are delivered after the subject has been diagnosed with thedisorder and before the disorder has been cured or eliminated. In someembodiments, the delivery of one treatment is still occurring when thedelivery of the second begins, so that there is overlap. This issometimes referred to herein as “simultaneous” or “concurrent delivery.”In other embodiments, the delivery of one treatment ends before thedelivery of the other treatment begins. In some embodiments of eithercase, the treatment is more effective because of combinedadministration. For example, the second treatment is more effective,e.g., an equivalent effect is seen with less of the second treatment, orthe second treatment reduces symptoms to a greater extent, than would beseen if the second treatment were administered in the absence of thefirst treatment, or the analogous situation is seen with the firsttreatment. In some embodiments, delivery is such that the reduction in asymptom, or other parameter related to the disorder is greater than whatwould be observed with one treatment delivered in the absence of theother. The effect of the two treatments can be partially additive,wholly additive, or greater than additive. The delivery can be such thatan effect of the first treatment delivered is still detectable when thesecond is delivered.

Anti-TIM-3 antibody molecules can be administered in combination withone or more of the existing modalities for treating cancers, including,but not limited to: surgery; radiation therapy (e.g., external-beamtherapy which involves three dimensional, conformal radiation therapywhere the field of radiation is designed.

In certain aspects, the anti-TIM-3 antibody is co-administered with asecond agent that acts on TIM-3 or another element of a TIM-3 pathway.

In some embodiments, e.g., when treating infectious disease, theanti-TIM-3 antibody may be co-administered with, e.g., an antibiotic, ananti-viral agent, or an anti-fungal agent.

In some embodiments, e.g., when treating Crohn's disease, the anti-TIM-3antibody may be co-administered with, e.g. an anti-inflammatory drugsuch as 5-aminosalicylic acid (5-ASA), prednisone, or hydrocortisone;purine analogs such as azathioprine; antimetabolites such asmethotrexate; TNF-alpha inhibitors, e.g., a monoclonal antibody to tumornecrosis factor alpha (TNF-α), e.g., infliximab, adalimumab, orcertolizumab; or integrin inhibitors, e.g., a monoclonal antibody toalpha-4-integrin, e.g., natalizumab.

In some embodiments, e.g., when treating multiple sclerosis, theanti-TIM-3 antibody may be co-administered with, e.g. an interferon suchas interferon beta-1a, interferon beta-1b, an interferon analog, arandom amino acid polymer such as glatiramer acetate; a type IItopoisomerase inhibitor such as mitoxantrone; an integrin inhibitor,e.g., a monoclonal antibody to alpha-4-integrin, e.g., natalizumab; asphingosine 1-phosphate receptor modulator, e.g., fingolimod; apyrimidines synthesis inhibitor, e.g., a dihydroorotate dehydrogenaseinhibitor such as teriflunomide; and other immunomodulatory agents suchas dimethyl fumarate.

In some embodiments, e.g., when treating sepsis, the anti-TIM-3 antibodymay be co-administered with, e.g. antibiotics; vasopressors such asnorepinephrine or dopamine; steroids; Recombinant activated protein C(drotrecogin alpha); intravenous fluids; and ventilation.

In some embodiments, e.g., when treating SIRS (Systemic InflammatoryResponse Syndrome) the anti-TIM-3 antibody may be co-administered with,e.g. antibiotics; steroids; antioxidants; or intravenous fluids.

In some embodiments, e.g., when treating glomerulonephritis, theanti-TIM-3 antibody may be co-administered with, e.g., steroids; analkylating agent such as cyclophosphamide; or a purine analog such asazathioprine.

Combinations of TIM-3 antibody molecules with one or more secondtherapeutics are provided herein. Many of the combinations in thissection are useful in treating cancer, but other indications are alsodescribed. This section focuses on combinations of anti-TIM-3 antibodymolecules, optionally in combination with one or more immunomodulators(e.g., an anti-PD-1 antibody molecule, an anti-LAG-3 antibody molecule,or an anti-PD-L1 antibody molecule), with one or more of the agentsdescribed in Table 6. In the combinations herein below, in oneembodiment, the anti-TIM-3 antibody molecule comprises (i) a heavy chainvariable region (VH) comprising a VHCDR1 amino acid sequence chosen fromSEQ ID NO: 3 or SEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQ ID NO:4, SEQ ID NO: 10, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 30, or SEQ IDNO: 31; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and (ii) alight chain variable region (VL) comprising a VLCDR1 amino acid sequenceof SEQ ID NO: 6 or SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ IDNO: 7 or SEQ ID NO: 13, and a VLCDR3 amino acid sequence of SEQ ID NO: 8or SEQ ID NO: 14.

In one embodiment, the anti-TIM-3 antibody molecule, e.g., an anti-TIM-3antibody molecule as described herein, alone or in combination with oneor more other immunomodulators, is used in combination with a PKCinhibitor, Sotrastaurin (Compound A1), or a compound disclosed in PCTPublication No. WO 2005/039549, to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the PKC inhibitor is disclosed inTable 6, or in a publication recited in Table 6, e.g., in the A1 row ofTable 6. In one embodiment, the PKC inhibitor is Sotrastaurin (CompoundA1) or a compound disclosed in PCT Publication No. WO 2005/039549. Inone embodiment, a TIM-3 antibody molecule is used in combination withSotrastaurin (Compound A1), or a compound as described in PCTPublication No. WO 2005/039549, to treat a disorder such as a cancer, amelanoma, a non-Hodgkin lymphoma, an inflammatory bowel disease,transplant rejection, an ophthalmic disorder, or psoriasis.

In certain embodiments, Sotrastaurin (Compound A1) is administered at adose of about 20 to 600 mg, e.g., about 200 to about 600 mg, about 50 mgto about 450 mg, about 100 mg to 400 mg, about 150 mg to 350 mg, orabout 200 mg to 300 mg, e.g., about 50 mg, 100 mg, 150 mg, 200 mg, 300mg, 400 mg, 500 mg, or 600 mg. The dosing schedule can vary from e.g.,every other day to daily, twice or three times a day.

In one embodiment, the anti-TIM-3 antibody molecule, e.g., an anti-TIM-3antibody molecule as described herein, alone or in combination with oneor more other immunomodulators, is used in combination with a BCR-ABLinhibitor, TASIGNA (Compound A2, or a compound disclosed in PCTPublication No. WO 2004/005281, to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the BCR-ABL inhibitor is TASIGNA,or a compound disclosed in PCT Publication No. WO 2004/005281. In oneembodiment, a TIM-3 antibody molecule is used in combination withTASIGNA (Compound A2), or a compound as described in PCT Publication No.WO 2004/005281, to treat a disorder such as a lymphocytic leukemia,Parkinson's Disease, a neurologic cancer, a melanoma, adigestive/gastrointestinal cancer, a colorectal cancer, a myeloidleukemia, a head and neck cancer, or pulmonary hypertension.

In one embodiment, the BCR-ABL inhibitor or TASIGNA is administered at adose of about 300 mg (e.g., twice daily, e.g., for newly diagnosed Ph+CML-CP), or about 400 mg, e.g., twice daily, e.g., for resistant orintolerant Ph+ CML-CP and CML-AP). BCR-ABL inhibitor or a Compound A2 isadministered at a dose of about 300-400 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an HSP90 inhibitor, such as5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide(Compound A3), or a compound disclosed in PCT Publication No. WO2010/060937 or WO 2004/072051, to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the HSP90 inhibitor is5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide(Compound A3), or a compound disclosed in PCT Publication No. WO2010/060937 or WO 2004/072051. In one embodiment, a TIM-3 antibodymolecule is used in combination with5-(2,4-dihydroxy-5-isopropylphenyl)-N-ethyl-4-(4-(morpholinomethyl)phenyl)isoxazole-3-carboxamide(Compound A3), or a compound as described in PCT Publication No. WO2010/060937 or WO 2004/072051, to treat a disorder such as a cancer, amultiple myeloma, a non-small cell lung cancer, a lymphoma, a gastriccancer, a breast cancer, a digestive/gastrointestinal cancer, apancreatic cancer, a colorectal cancer, a solid tumor, or ahematopoiesis disorder.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an inhibitor of PI3K and/or mTOR, Dactolisib (CompoundA4) or8-(6-Methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one(Compound A41), or a compound disclosed in PCT Publication No. WO2006/122806, to treat a disorder, e.g., a disorder described herein. Inone embodiment, the PI3K and/or mTOR inhibitor is Dactolisib (CompoundA4),8-(6-Methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one(Compound A41), or a compound disclosed in PCT Publication No. WO2006/122806. In one embodiment, a TIM-3 antibody molecule is used incombination with Dactolisib (Compound A4),8-(6-Methoxy-pyridin-3-yl)-3-methyl-1-(4-piperazin-1-yl-3-trifluoromethyl-phenyl)-1,3-dihydro-imidazo[4,5-c]quinolin-2-one(Compound A41), or a compound described in PCT Publication No. WO2006/122806, to treat a disorder such as a cancer, a prostate cancer, aleukemia (e.g., lymphocytic leukemia), a breast cancer, a brain cancer,a bladder cancer, a pancreatic cancer, a renal cancer, a solid tumor, ora liver cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an FGFR inhibitor,3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1-methylurea(Compound A5) or a compound disclosed in U.S. Pat. No. 8,552,002, totreat a disorder, e.g., a disorder described herein. In one embodiment,the FGFR inhibitor is3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1-methylurea(Compound A5) or a compound disclosed in U.S. Pat. No. 8,552,002. In oneembodiment, a TIM-3 antibody molecule is used in combination withCompound A5, or a compound as described in U.S. Pat. No. 8,552,002, totreat a disorder such as a digestive/gastrointestinal cancer, ahematological cancer, or a solid tumor.

In one embodiment, the FGFR inhibitor or3-(2,6-dichloro-3,5-dimethoxyphenyl)-1-(6-((4-(4-ethylpiperazin-1-yl)phenyl)amino)pyrimidin-4-yl)-1-methylurea(Compound A5) is administered at a dose of about 100-125 mg (e.g., perday), e.g., about 100 mg or about 125 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a PI3K inhibitor, Buparlisib (Compound A6), or acompound disclosed in PCT Publication No. WO 2007/084786, to treat adisorder, e.g., a disorder described herein. In one embodiment, the PI3Kinhibitor is Buparlisib (Compound A6) or a compound disclosed in PCTPublication No. WO 2007/084786. In one embodiment, a TIM-3 antibodymolecule is used in combination with Buparlisib (Compound A6), or acompound disclosed in PCT Publication No. WO 2007/084786, to treat adisorder such as, a prostate cancer, a non-small cell lung cancer, anendocrine cancer, a leukemia, an ovarian cancer, a melanoma, a bladdercancer, a breast cancer, a female reproductive system cancer, adigestive/gastrointestinal cancer, a colorectal cancer, a glioblastomamultiforme, a solid tumor, a non-Hodgkin lymphoma, a hematopoiesisdisorder, or a head and neck cancer.

In one embodiment, the PI3K inhibitor or Buparlisib (Compound A6) isadministered at a dose of about 100 mg (e.g., per day).

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an FGFR inhibitor,8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxaline-5-carboxamide(Compound A7) or a compound disclosed in PCT Publication No. WO2009/141386 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the FGFR inhibitor is8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxaline-5-carboxamide(CompoundA7) or a compound disclosed in a PCT Publication No. WO 2009/141386. Inone embodiment, the FGFR inhibitor is8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxaline-5-carboxamide(CompoundA7). In one embodiment, a TIM-3 antibody molecule is used in combinationwith8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxaline-5-carboxamide(CompoundA7), or a compound disclosed in PCT Publication No. WO 2009/141386, totreat a disorder such as a cancer characterized by angiogenesis.

In one embodiment, the FGFR inhibitor or8-(2,6-difluoro-3,5-dimethoxyphenyl)-N-(4-((dimethylamino)methyl)-1H-imidazol-2-yl)quinoxaline-5-carboxamide(Compound A7) is administered at a dose of e.g., from approximately 3 mgto approximately 5 g, more preferably from approximately 10 mg toapproximately 1.5 g per person per day, optionally divided into 1 to 3single doses which may, for example, be of the same size.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a PI3K inhibitor,(S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide(Compound A8) or a compound disclosed PCT Publication No. WO 2010/029082to treat a disorder, e.g., a disorder described herein. In oneembodiment, the PI3K inhibitor is(S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide(Compound A8) or a compound disclosed PCT Publication No. WO2010/029082. In one embodiment, a TIM-3 antibody molecule is used incombination with(S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide(Compound A8), or a compound disclosed PCT Publication No. WO2010/029082, to treat a disorder such as a gastric cancer, a breastcancer, a pancreatic cancer, a digestive/gastrointestinal cancer, asolid tumor, and a head and neck cancer.

In one embodiment, the PI3K inhibitor or(S)-N1-(4-methyl-5-(2-(1,1,1-trifluoro-2-methylpropan-2-yl)pyridin-4-yl)thiazol-2-yl)pyrrolidine-1,2-dicarboxamide(Compound A8) is administered at a dose of about 150-300, 200-300,200-400, or 300-400 mg (e.g., per day), e.g., about 200, 300, or 400 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an inhibitor of cytochrome P450 (e.g., a CYP17inhibitor) or a compound disclosed in PCT Publication No. WO2010/149755, to treat a disorder, e.g., a disorder described herein. Inone embodiment, the cytochrome P450 inhibitor (e.g., the CYP17inhibitor) is a compound disclosed in PCT Publication No. WO2010/149755. In one embodiment, a TIM-3 antibody molecule is used incombination with a compound disclosed in PCT Publication No. WO2010/149755, to treat prostate cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an HDM2 inhibitor,(S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one(CompoundA10) or a compound disclosed in PCT Publication No. WO 2011/076786 totreat a disorder, e.g., a disorder described herein). In one embodiment,the HDM2 inhibitor is(S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one(Compound A10) or a compound disclosed in PCT Publication No. WO2011/076786. In one embodiment, a TIM-3 antibody molecule is used incombination with(S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one(Compound A10), or a compound disclosed in PCTPublication No. WO2011/076786, to treat a disorder such as a solid tumor.

In one embodiment, the HDM2 inhibitor or(S)-1-(4-chlorophenyl)-7-isopropoxy-6-methoxy-2-(4-(methyl(((1r,4S)-4-(4-methyl-3-oxopiperazin-1-yl)cyclohexyl)methyl)amino)phenyl)-1,2-dihydroisoquinolin-3(4H)-one(Compound A10) is administered at a dose of about 400 to 700 mg, e.g.,administered three times weekly, 2 weeks on and one week off. In someembodiments, the dose is about 400, 500, 600, or 700 mg; about 400-500,500-600, or 600-700 mg, e.g., administered three times weekly.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an iron chelating agent, Deferasirox (also known asEXJADE; Compound A11), or a compound disclosed in PCT Publication No. WO1997/049395 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the iron chelating agent is Deferasirox or a compounddisclosed in PCT Publication No. WO 1997/049395. In one embodiment, theiron chelating agent is Deferasirox (Compound A11). In one embodiment, aTIM-3 antibody molecule is used in combination with Deferasirox(Compound A11), or a compound disclosed in PCT Publication No. WO1997/049395, to treat iron overload, hemochromatosis, or myelodysplasia.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an aromatase inhibitor, Letrozole (also known asFEMARA; Compound A12), or a compound disclosed in U.S. Pat. No.4,978,672 to treat a disorder, e.g., a disorder described herein. In oneembodiment, the aromatase inhibitor is Letrozole (Compound A12) or acompound disclosed in U.S. Pat. No. 4,978,672. In one embodiment, aTIM-3 antibody molecule is used in combination with Letrozole (CompoundA12), or a compound disclosed in U.S. Pat. No. 4,978,672, to treat adisorder such as a cancer, a leiomyosarcoma, an endometrium cancer, abreast cancer, a female reproductive system cancer, or a hormonedeficiency.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a PI3K inhibitor, e.g., a pan-PI3K inhibitor,(4S,5R)-3-(2′-amino-2-morpholino-4′-(trifluoromethyl)-[4,5′-bipyrimidin]-6-yl)-4-(hydroxymethyl)-5-methyloxazolidin-2-one(Compound A13) or a compound disclosed in PCT Publication No.WO2013/124826 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the PI3K inhibitor is(4S,5R)-3-(2′-amino-2-morpholino-4′-(trifluoromethyl)-[4,5′-bipyrimidin]-6-yl)-4-(hydroxymethyl)-5-methyloxazolidin-2-one(Compound A13) or a compound disclosed in PCT Publication No.WO2013/124826. In one embodiment, a TIM-3 antibody molecule is used incombination with(4S,5R)-3-(2′-amino-2-morpholino-4′-(trifluoromethyl)-[4,5′-bipyrimidin]-6-yl)-4-(hydroxymethyl)-5-methyloxazolidin-2-one(Compound A13), or a compound disclosed in PCT Publication No.WO2013/124826, to treat a disorder such as a cancer or an advanced solidtumor.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an inhibitor of p53 and/or a p53/Mdm2 interaction,(S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one(Compound A14), or a compound disclosed in PCT Publication No.WO2013/111105 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the p53 and/or a p53/Mdm2 interaction inhibitor is(S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one(Compound A14) or a compound disclosed in PCT Publication No.WO2013/111105. In one embodiment, a TIM-3 antibody molecule is used incombination with(S)-5-(5-chloro-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)-6-(4-chlorophenyl)-2-(2,4-dimethoxypyrimidin-5-yl)-1-isopropyl-5,6-dihydropyrrolo[3,4-d]imidazol-4(1H)-one(Compound A14), or a compound disclosed in PCT Publication No.WO2013/111105, to treat a disorder such as a cancer or a soft tissuesarcoma.

In another embodiment, anti-TIM-3 antibody molecule, e.g., an anti-TIM-3antibody molecule as described herein, alone or in combination with oneor more other immunomodulators, is used in combination with a CSF-1Rtyrosine kinase inhibitor,4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide(Compound A15), or a compound disclosed in PCT Publication No. WO2005/073224 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the CSF-1R tyrosine kinase inhibitor is4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide(Compound A15) or a compound disclosed in PCT Publication No. WO2005/073224. In one embodiment, anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, is used in combinationwith4-((2-(((1R,2R)-2-hydroxycyclohexyl)amino)benzo[d]thiazol-6-yl)oxy)-N-methylpicolinamide(Compound A15) or a compound disclosed in PCT Publication No. WO2005/073224, to treat a disorder such as cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an apoptosis inducer and/or an angiogenesis inhibitor,such as Imatinib mesylate (also known as GLEEVEC; Compound A16) or acompound disclosed in PCT Publication No. WO1999/003854 to treat adisorder, e.g., a disorder described. In one embodiment, the apoptosisinducer and/or an angiogenesis inhibitor is Imatinib mesylate (CompoundA16) or a compound disclosed in PCT Publication No. WO1999/003854. Inone embodiment, a TIM-3 antibody molecule is used in combination withImatinib mesylate (Compound A16), or a compound disclosed in PCTPublication No. WO1999/003854, to treat a disorder such as a cancer, amultiple myeloma, a prostate cancer, a non-small cell lung cancer, alymphoma, a gastric cancer, a melanoma, a breast cancer, a pancreaticcancer, a digestive/gastrointestinal cancer, a colorectal cancer, aglioblastoma multiforme, a liver cancer, a head and neck cancer, asthma,multiple sclerosis, allergy, Alzheimer's dementia, amyotrophic lateralsclerosis, or rheumatoid arthritis.

In certain embodiments, Imatinib mesylate (Compound A16) is administeredat a dose of about 100 to 1000 mg, e.g., about 200 mg to 800 mg, about300 mg to 700 mg, or about 400 mg to 600 mg, e.g., about 200 mg, 300 mg,400 mg, 500 mg, 600 mg, or 700 mg. The dosing schedule can vary frome.g., every other day to daily, twice or three times a day. In oneembodiment, Imatinib mesylate is administered at an oral dose from about100 mg to 600 mg daily, e.g., about 100 mg, 200 mg, 260 mg, 300 mg, 400mg, or 600 mg daily.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a JAK inhibitor,2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide(Compound A17), or a dihydrochloric salt thereof, or a compounddisclosed in PCT Publication No. WO 2007/070514, to treat a disorder,e.g., a disorder described herein. In one embodiment, the JAK inhibitoris2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide(Compound A17), or a dihydrochloric salt thereof, or a compounddisclosed in PCT Publication No. WO 2007/070514. In one embodiment, aTIM-3 antibody molecule is used in combination with2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide(Compound A17), or a dihydrochloric salt thereof, or a compounddisclosed in PCT Publication No. WO 2007/070514, to treat a disordersuch as colorectal cancer, myeloid leukemia, hematological cancer,autoimmune disease, non-Hodgkin lymphoma, or thrombocythemia.

In one embodiment, the JAK inhibitor or a2-fluoro-N-methyl-4-(7-(quinolin-6-ylmethyl)imidazo[1,2-b][1,2,4]triazin-2-yl)benzamide(Compound A17), or a dihydrochloric salt thereof is administered at adose of about 400-600 mg (e.g., per day), e.g., about 400, 500, or 600mg, or about 400-500 or 500-600 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a JAK inhibitor, Ruxolitinib Phosphate (also known asJAKAFI; Compound A18) or a compound disclosed in PCT Publication No. WO2007/070514 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the JAK inhibitor is Ruxolitinib Phosphate (CompoundA18) or a compound disclosed in PCT Publication No. WO 2007/070514. Inone embodiment, a TIM-3 antibody molecule is used in combination withRuxolitinib Phosphate (Compound A18), or a compound disclosed in PCTPublication No. WO 2007/070514, to treat a disorder such as a prostatecancer, a lymphocytic leukemia, a multiple myeloma, a lymphoma, a lungcancer, a leukemia, cachexia, a breast cancer, a pancreatic cancer,rheumatoid arthritis, psoriasis, a colorectal cancer, a myeloidleukemia, a hematological cancer, an autoimmune disease, a non-Hodgkinlymphoma, or thrombocythemia.

In one embodiment, the JAK inhibitor or Ruxolitinib Phosphate (CompoundA18) is administered at a dose of about 15-25 mg, e.g., twice daily. Insome embodiments, the dose is about 15, 20, or 25 mg, or about 15-20 or20-25 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a deacetylase (DAC) inhibitor, Panobinostat (CompoundA19), or a compound disclosed in PCT Publication No. WO 2014/072493 totreat a disorder, e.g., a disorder described herein. In one embodiment,the DAC inhibitor is Panobinostat (Compound A19) or a compound disclosedin PCT Publication No. WO 2014/072493. In one embodiment, a TIM-3antibody molecule is used in combination with Panobinostat (CompoundA19), a compound disclosed in PCT Publication No. WO 2014/072493, totreat a disorder such as a small cell lung cancer, arespiratory/thoracic cancer, a prostate cancer, a multiple myeloma,myelodysplastic syndrome, a bone cancer, a non-small cell lung cancer,an endocrine cancer, a lymphoma, a neurologic cancer, a leukemia,HIV/AIDS, an immune disorder, transplant rejection, a gastric cancer, amelanoma, a breast cancer, a pancreatic cancer, a colorectal cancer, aglioblastoma multiforme, a myeloid leukemia, a hematological cancer, arenal cancer, a non-Hodgkin lymphoma, a head and neck cancer, ahematopoiesis disorders, or a liver cancer.

In one embodiment, the DAC inhibitor or Panobinostat (Compound A19) isadministered at a dose of about 20 mg (e.g., per day).

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an inhibitor of one or more of cytochrome P450 (e.g.,11B2), aldosterone or angiogenesis, Osilodrostat (Compound A20), or acompound disclosed in PCT Publication No. WO2007/024945 to treat adisorder, e.g., a disorder described herein. In one embodiment, theinhibitor of one or more of cytochrome P450 (e.g., 11B2), aldosterone orangiogenesis is Osilodrostat (Compound A20) or a compound disclosed inPCT Publication No. WO2007/024945. In one embodiment, a TIM-3 antibodymolecule is used in combination with Osilodrostat (Compound A20), or acompound disclosed in PCT Publication No. WO2007/024945, to treat adisorder such as Cushing's syndrome, hypertension, or heart failuretherapy.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a IAP inhibitor,(S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide(Compound A21) or a compound disclosed in U.S. Pat. No. 8,552,003 totreat a disorder, e.g., a disorder described herein. In one embodiment,the IAP inhibitor is(S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide(Compound A21) or a compound disclosed in U.S. Pat. No. 8,552,003. Inone embodiment, a TIM-3 antibody molecule is used in combination with(S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide(Compound A21), or a compound disclosed in U.S. Pat. No. 8,552,003, totreat a disorder such as a multiple myeloma, a breast cancer, an ovariancancer, a pancreatic cancer, or a hematopoiesis disorder.

In one embodiment, the IAP inhibitor or(S)-N-((S)-1-cyclohexyl-2-((S)-2-(4-(4-fluorobenzoyl)thiazol-2-yl)pyrrolidin-1-yl)-2-oxoethyl)-2-(methylamino)propanamide(Compound A21) or a compound disclosed in U.S. Pat. No. 8,552,003 isadministered at a dose of approximately 1800 mg, e.g., once weekly.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a Smoothened (SMO) inhibitor, Sonidegib phosphate (CompoundA22),(R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1-yl)pyrazin-2-yl)propan-2-ol(Compound A25), or a compound disclosed in PCT Publication No. WO2007/131201 or WO 2010/007120 to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the SMO inhibitor is Sonidegibphosphate (Compound A22),(R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1-yl)pyrazin-2-yl)propan-2-ol(Compound A25), or a compound disclosed in PCT Publication No. WO2007/131201 or WO 2010/007120. In one embodiment, a TIM-3 antibodymolecule is used in combination with Sonidegib phosphate (Compound A22),(R)-2-(5-(4-(6-benzyl-4,5-dimethylpyridazin-3-yl)-2-methylpiperazin-1-yl)pyrazin-2-yl)propan-2-ol(Compound A25), or a compound disclosed in PCT Publication No. WO2007/131201 or WO 2010/007120 to treat a disorder such as a cancer, amedulloblastoma, a small cell lung cancer, a prostate cancer, a basalcell carcinoma, a pancreatic cancer, or an inflammation.

In certain embodiments, Sonidegib phosphate (Compound A22) isadministered at a dose of about 20 to 500 mg, e.g., about 40 mg to 400mg, about 50 mg to 300 mg, or about 100 mg to 200 mg, e.g., about 50 mg,100 mg, 150 mg, 200 mg, 250 mg, or 300 mg. The dosing schedule can varyfrom e.g., every other day to daily, twice or three times a day.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an Alk inhibitor, ceritinib (also known as ZYKADIA;Compound A23) or a compound disclosed in PCT Publication No. WO2007/131201 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the Alk inhibitor is ceritinib (Compound A23) or acompound disclosed in PCT Publication No. WO 2007/131201. In oneembodiment, a TIM-3 antibody molecule is used in combination withceritinib (Compound A23), or a compound disclosed in PCT Publication No.WO 2007/131201, to treat a disorder such as non-small cell lung canceror solid tumors.

In one embodiment, the Alk inhibitor or ceritinib (Compound A23) isadministered at a dose of approximately 750 mg, e.g., once daily.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a JAK and/or CDK4/6 inhibitor,7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A24), or a compound disclosed in U.S. Pat. No. 8,415,355 orU.S. Pat. No. 8,685,980 to treat a disorder, e.g., a disorder describedherein. In one embodiment, the JAK and/or CDK4/6 inhibitor is7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A24) or a compound disclosed in U.S. Pat. No. 8,415,355 orU.S. Pat. No. 8,685,980. In one embodiment, a TIM-3 antibody molecule isused in combination with7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A24), or a compound disclosed in U.S. Pat. No. 8,415,355 orU.S. Pat. No. 8,685,980, to treat a disorder such as a lymphoma, aneurologic cancer, a melanoma, a breast cancer, or a solid tumor.

In one embodiment, the JAK and/or CDK4/6 inhibitor or7-cyclopentyl-N,N-dimethyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A24) is administered at a dose of approximately 200-600 mg,e.g., per day. In one embodiment, the compound is administered at a doseof about 200, 300, 400, 500, or 600 mg, or about 200-300, 300-400,400-500, or 500-600 mg.

In another embodiment, the antibody molecule, e.g., an anti-TIM-3antibody molecule as described herein, alone or in combination with oneor more other immunomodulators, is used in combination a prolactinreceptor (PRLR) inhibitor, a human monoclonal antibody molecule(Compound A26) as disclosed in U.S. Pat. No. 7,867,493), to treat adisorder, e.g., a disorder described herein. In one embodiment, the PRLRinhibitor is a human monoclonal antibody (Compound A26) disclosed inU.S. Pat. No. 7,867,493. In one embodiment, a TIM-3 antibody molecule isused in combination with human monoclonal antibody molecule (CompoundA26) described in U.S. Pat. No. 7,867,493 to treat a disorder such as, acancer, a prostate cancer, or a breast cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a PIM Kinase inhibitor,N-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide(Compound A27) or a compound disclosed in PCT Publication No. WO2010/026124 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the PIM Kinase inhibitor isN-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide(Compound A27) or a compound disclosed in PCT Publication No. WO2010/026124. In one embodiment, a TIM-3 antibody molecule is used incombination withN-(4-((1R,3S,5S)-3-amino-5-methylcyclohexyl)pyridin-3-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide(Compound A27), or a compound disclosed in PCT Publication No. WO2010/026124, to treat a disorder such as a multiple myeloma,myelodysplastic syndrome, a myeloid leukemia, or a non-Hodgkin lymphoma.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a Wnt signaling inhibitor,2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28) or a compound disclosed in PCT publication No. WO2010/101849 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the Wnt signaling inhibitor is2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28) or a compound disclosed in PCT publication No. WO2010/101849. In one embodiment, the Wnt signaling inhibitor is2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28). In one embodiment, a TIM-3 antibody molecule is used incombination with2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28), or a compound disclosed in PCT publication No. WO2010/101849, to treat a disorder such as a solid tumor (e.g., a head andneck cancer, a squamous cell carcinoma, a breast cancer, a pancreaticcancer, or a colon cancer).

In certain embodiments,2-(2′,3-dimethyl-[2,4′-bipyridin]-5-yl)-N-(5-(pyrazin-2-yl)pyridin-2-yl)acetamide(Compound A28) is administered at a dose of about 1 to 50 mg, e.g.,about 2 mg to 45 mg, about 3 mg to 40 mg, about 5 mg to 35 mg, 5 mg to10 mg, or about 10 mg to 30 mg, e.g., about 2 mg, 5 mg, 10 mg, 20 mg, 30mg, or 40 mg. The dosing schedule can vary from e.g., every other day todaily, twice or three times a day.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a BRAF inhibitor, Encorafenib (Compound A29), or acompound disclosed in PCT Publication No. WO 2011/025927 to treat adisorder, e.g., a disorder described herein. In one embodiment, the BRAFinhibitor is Encorafenib (Compound A29) or a compound disclosed in PCTPublication No. WO 2011/025927. In one embodiment, a TIM-3 antibodymolecule is used in combination with Encorafenib (Compound A29), or acompound disclosed in PCT Publication No. WO 2011/025927, to treat adisorder such as a non-small cell lung cancer, a melanoma, or acolorectal cancer.

In one embodiment, the BRAF inhibitor or Encorafenib (Compound A29) isadministered at a dose of about 200-300, 200-400, or 300-400 mg, e.g.,per day. In one embodiment, the compound is administered at a dose ofabout 200, about 300 or about 400 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a CDK4/6 inhibitor,7-cyclopentyl-N,N-dimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A30), or a compound disclosed in PCT publication No. WO2011/101409 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the CDK4/6 inhibitor is7-cyclopentyl-N,N-dimethyl-2-((5-((1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A30) or a compound disclosed in PCT publication No. WO2011/101409. In one embodiment, a TIM-3 antibody molecule is used incombination with7-cyclopentyl-N,N-dimethyl-2-((5(1R,6S)-9-methyl-4-oxo-3,9-diazabicyclo[4.2.1]nonan-3-yl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxamide(Compound A30), or a compound disclosed in PCT publication No. WO2011/101409, to treat a disorder such as a cancer, a mantle celllymphoma, a liposarcoma, a non-small cell lung cancer, a melanoma, asquamous cell esophageal cancer, or a breast cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a HER3 inhibitor, Compound A31, or a compound disclosedin PCT Publication No. WO 2012/022814, to treat a disorder, e.g., adisorder described herein. In one embodiment, the HER3 inhibitor isCompound A31 or a compound disclosed in PCT Publication WO 2012/022814.In one embodiment, a TIM-3 antibody molecule is used in combination withCompound A31, or a compound disclosed in PCT Publication WO 2012/022814,to treat a disorder such as a gastric cancer, an esophageal cancer, ahead and neck cancer, a squamous cell carcinoma, a stomach cancer, abreast cancer (e.g., metastatic breast cancer), or adigestive/gastrointestinal cancer.

In some embodiments, Compound A31 is a human monoclonal antibodymolecule.

In one embodiment, the HER3 inhibitor or Compound A31 is administered ata dose of about 3, 10, 20, or 40 mg/kg, e.g., once weekly (QW). In oneembodiment, the compound is administered at a dose of about 3-10, 10-20,or 20-40 mg/kg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an FGFR2 and/or FGFR4 inhibitor, Compound A32, or a compounddisclosed in a publication PCT Publication No. WO 2014/160160 (e.g., anantibody molecule drug conjugate against an FGFR2 and/or FGFR4, e.g.,mAb 12425), to treat a disorder, e.g., a disorder described herein. Inone embodiment, the FGFR2 and/or FGFR4 inhibitor is Compound A32 or acompound disclosed in a publication PCT Publication No. WO 2014/160160.In one embodiment, a TIM-3 antibody molecule is used in combination withCompound A32, or a compound as described in Table 6, to treat a disordersuch as a cancer, a gastric cancer, a breast cancer, a rhabdomyosarcoma,a liver cancer, an adrenal cancer, a lung cancer, an esophageal cancer,a colon cancer, or an endometrial cancer.

In some embodiments, Compound A32 is an antibody molecule drug conjugateagainst an FGFR2 and/or FGFR4, e.g., mAb 12425.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an M-CSF inhibitor, Compound A33, or a compound disclosed inPCT Publication No. WO 2004/045532 (e.g., an antibody molecule or Fabfragment against M-CSF), to treat a disorder, e.g., a disorder describedherein. In one embodiment, the M-CSF inhibitor is Compound A33 or acompound disclosed in PCT Publication No. WO 2004/045532. In oneembodiment, a TIM-3 antibody molecule is used in combination withCompound A33, or a compound as described in PCT Publication No. WO2004/045532, to treat a disorder such as a cancer, a prostate cancer, abreast cancer, or pigmented villonodular synovitis (PVNS).

In embodiments, Compound A33 is a monoclonal antibody molecule againstM-CSF or a fragment (e.g., Fab fragment) thereof. In embodiments, theM-CSF inhibitor or Compound A33 is administered at an average dose ofabout 10 mg/kg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a MEK inhibitor, Binimetinib (Compound A34), or acompound disclosed in PCT Publication No. WO 2003/077914 to treat adisorder, e.g., a disorder described herein. In one embodiment, the MEKinhibitor is Binimetinib (Compound A34), or a compound disclosed in PCTPublication No. WO 2003/077914. In one embodiment, a TIM-3 antibodymolecule is used in combination with Binimetinib (Compound A34), or acompound disclosed in PCT Publication No. WO 2003/077914, to treat adisorder such as a non-small cell lung cancer, a multisystem geneticdisorder, a melanoma, an ovarian cancer, a digestive/gastrointestinalcancer, a rheumatoid arthritis, or a colorectal cancer.

In one embodiment, the MEK inhibitor or Binimetinib (Compound A34) isadministered at a dose of about 45 mg, e.g., twice daily.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an inhibitor of one or more of c-KIT, histamine release,Flt3 (e.g., FLK2/STK1) or PKC, Midostaurin (Compound A35) or a compounddisclosed in PCT Publication No. WO 2003/037347 to treat a disorder,e.g., a disorder described herein. In one embodiment, the inhibitor isMidostaurin (Compound A35) or compound disclosed in PCT Publication No.WO 2003/037347. In one embodiment, the inhibitor of one or more ofc-KIT, histamine release, Flt3 (e.g., FLK2/STK1) or PKC is Midostaurin.In one embodiment, a TIM-3 antibody molecule is used in combination withMidostaurin (Compound A35), or compound disclosed in PCT Publication No.WO 2003/037347, to treat a disorder such as a cancer, a colorectalcancer, a myeloid leukemia, myelodysplastic syndrome, an age-relatedmascular degeration, a diabetic complication, or a dermatologicdisorder.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a TOR inhibitor (e.g., mTOR inhibitor), Everolimus(also known as AFINITOR; Compound A36) or a Compound disclosed in PCTPublication No. WO 2014/085318 to treat a disorder, e.g., a disorderdescribed herein). In one embodiment, the TOR inhibitor is Everolimus(Compound A36) or a Compound disclosed in PCT Publication No. WO2014/085318. In one embodiment, a TIM-3 antibody molecule is used incombination with Everolimus (Compound A36) to treat a disorder such asan interstitial lung disease, a small cell lung cancer, arespiratory/thoracic cancer, a prostate cancer, a multiple myeloma, asarcoma, an age-related macular degeneration, a bone cancer, tuberoussclerosis, a non-small cell lung cancer, an endocrine cancer, alymphoma, a neurologic disorders, an astrocytoma, a cervical cancer, aneurologic cancer, a leukemia, an immune disorders, transplantrejection, a gastric cancer, a melanoma, epilepsy, a breast cancer, or abladder cancer.

In one embodiment, the TOR inhibitor or Everolimusis (Compound A36)administered at a dose of about 2.5-20 mg/day. In one embodiment, thecompound is administered at a dose of about 2.5, 5, 10, or 20 mg/day,e.g., about 2.5-5, 5-10, or 10-20 mg/day.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an inhibitor of one or more of VEGFR-2, PDGFRbeta, KIT orRaf kinase C,1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine(Compound A37) or a compound disclosed in PCT Publication No. WO2007/030377 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the inhibitor of one or more of VEGFR-2, PDGFRbeta, KITor Raf kinase C is1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine(Compound A37) or a compound disclosed in PCT Publication No. WO2007/030377. In one embodiment, a TIM-3 antibody molecule is used incombination with1-methyl-5-((2-(5-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-4-yl)oxy)-N-(4-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2-amine(Compound A37), or a compound disclosed in PCT Publication No. WO2007/030377, to treat a disorder such as a cancer, a melanoma, or asolid tumor.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a somatostatin agonist and/or growth hormone releaseinhibitor, Pasireotide diaspartate (also known as SIGNIFOR; CompoundA38) or a compound disclosed in PCT Publication No. WO2002/010192 orU.S. Pat. No. 7,473,761 to treat a disorder, e.g., a disorder describedherein. In one embodiment, the somatostatin agonist and/or growthhormone release inhibitor is Pasireotide diaspartate (Compound A38) or acompound disclosed in PCT Publication No. WO2002/010192 or U.S. Pat. No.7,473,761. In one embodiment, a TIM-3 antibody molecule is used incombination with Pasireotide diaspartate (Compound A38), or a compounddisclosed in PCT Publication No. WO2002/010192 or U.S. Pat. No.7,473,761, to treat a disorder such as a prostate cancer, an endocrinecancer, a nurologic cancer, a skin cancer (e.g., a melanoma), apancreatic cancer, a liver cancer, Cushing's syndrome, agastrointestinal disorder, acromegaly, a liver and biliary tractdisorder, or liver cirrhosis.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a signal transduction modulator and/or angiogenesisinhibitor, Dovitinib (Compound A39) or a compound disclosed in PCTPublication No. WO 2009/115562 to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the signal transduction modulatorand/or angiogenesis inhibitor is Dovitinib (Compound A39) or a compounddisclosed in PCT Publication No. WO 2009/115562. In one embodiment, aTIM-3 antibody molecule is used in combination with Dovitinib (CompoundA39), or a compound disclosed in PCT Publication No. WO 2009/115562, totreat a disorder such as a cancer, a respiratory/thoracic cancer, amultiple myeloma, a prostate cancer, a non-small cell lung cancer, anendocrine cancer, or a neurological genetic disorder.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an EGFR inhibitor,(R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide(Compound A40) or a compound disclosed in PCT Publication No. WO2013/184757 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the EGFR inhibitor is(R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide(Compound A40) or a compound disclosed in PCT Publication No. WO2013/184757. In one embodiment, a TIM-3 antibody molecule is used incombination with(R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide(Compound A40), or a compound disclosed in PCT Publication No. WO2013/184757, to treat a disorder such as a cancer, e.g., a solid tumor.

In one embodiment, the EGFR inhibitor or(R,E)-N-(7-chloro-1-(1-(4-(dimethylamino)but-2-enoyl)azepan-3-yl)-1H-benzo[d]imidazol-2-yl)-2-methylisonicotinamide(Compound A40) is administered at a dose of 150-250 mg, e.g., per day.In one embodiment, the compound is administered at a dose of about 150,200, or 250 mg, or about 150-200 or 200-250 mg.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an ALK inhibitor,N⁶-(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N⁴-(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidine-4,6-diamine(Compound A42) or a compound disclosed in PCT Publication No. WO2008/073687 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the ALK inhibitor isN⁶-(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N⁴-(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidine-4,6-diamine(Compound A42) or a compound disclosed in PCT Publication No. WO2008/073687. In one embodiment, a TIM-3 antibody molecule is used incombination withN⁶-(2-isopropoxy-5-methyl-4-(1-methylpiperidin-4-yl)phenyl)-N⁴-(2-(isopropylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidine-4,6-diamine(Compound A42), or a compound disclosed in PCT Publication No. WO2008/073687, to treat a disorder such as a cancer, an anaplasticlarge-cell lymphoma (ALCL), a non-small cell lung carcinoma (NSCLC), ora neuroblastoma.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination an IGF-1R inhibitor,3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thietane1,1-dioxide (Compound A43),5-chloro-N²-(2-fluoro-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A44), or5-chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A45) or a compound disclosed in PCT Publication No. WO2010/002655 to treat a disorder, e.g., a disorder described. In oneembodiment, the IGF-1R inhibitor is3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thietane1,1-dioxide (Compound A43),5-chloro-N²-(2-fluoro-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A44),5-chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A45), or a compound disclosed in PCT Publication No. WO2010/002655. In one embodiment, a TIM-3 antibody molecule is used incombination with3-(4-(4-((5-chloro-4-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)amino)-5-fluoro-2-methylphenyl)piperidin-1-yl)thietane1,1-dioxide (Compound A43),5-chloro-N²-(2-fluoro-5-methyl-4-(1-(tetrahydro-2H-pyran-4-yl)piperidin-4-yl)phenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A44),5-chloro-N2-(4-(1-ethylpiperidin-4-yl)-2-fluoro-5-methylphenyl)-N⁴-(5-methyl-1H-pyrazol-3-yl)pyrimidine-2,4-diamine(Compound A45), or a compound disclosed in PCT Publication No. WO2010/002655, to treat a disorder such as a cancer or a sarcoma.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination a P-Glycoprotein 1 inhibitor, Valspodar (also known asAMDRAY; Compound A46) or a compound disclosed in EP 296122 to treat adisorder, e.g., a disorder described herein. In one embodiment, theP-Glycoprotein 1 inhibitor is Valspodar (Compound A46) or a compounddisclosed in EP 296122. In one embodiment, a TIM-3 antibody molecule isused in combination with Valspodar (Compound A46), or a compounddisclosed in EP 296122, to treat a disorder such as a cancer or adrug-resistant tumor.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination one or more of a VEGFR inhibitor, Vatalanib succinate(Compound A47) or a compound disclosed in EP 296122 to treat a disorder,e.g., a disorder described herein. In one embodiment, the VEGFRinhibitor is Vatalanib succinate (Compound A47) or a compound disclosedin EP 296122. In one embodiment, a TIM-3 antibody molecule is used incombination with Vatalanib succinate (Compound A47), or a compounddisclosed in EP 296122, to treat cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an IDH inhibitor or a compound disclosed inWO2014/141104 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the IDH inhibitor is a compound disclosed in PCTPublication No. WO2014/141104. In one embodiment, a TIM-3 antibodymolecule is used in combination with a compound disclosed inWO2014/141104 to treat a disorder such as a cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a BCL-ABL inhibitor or a compound disclosed in PCTPublication No. WO2013/171639, WO2013/171640, WO2013/171641, orWO2013/171642 to treat a disorder, e.g., a disorder described herein. Inone embodiment, the BCL-ABL inhibitor is a compound disclosed in PCTPublication No. WO2013/171639, WO2013/171640, WO2013/171641, orWO2013/171642. In one embodiment, a TIM-3 antibody molecule is used incombination with a compound disclosed in PCT Publication No.WO2013/171639, WO2013/171640, WO2013/171641, or WO2013/171642 to treat adisorder such as a cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with a c-RAF inhibitor or a compound disclosed in PCTPublication No. WO2014/151616 to treat a disorder, e.g., a disorderdescribed herein. In one embodiment, the c-RAF inhibitor is Compound A50or a compound disclosed in PCT Publication No. WO2014/151616. In oneembodiment, a TIM-3 antibody molecule is used in combination with acompound disclosed in PCT Publication No. WO2014/151616 to treat adisorder such as a cancer.

In another embodiment, the anti-TIM-3 antibody molecule, e.g., ananti-TIM-3 antibody molecule as described herein, alone or incombination with one or more other immunomodulators, is used incombination with an ERK1/2 ATP competitive inhibitor or a compounddisclosed in International Patent Application No. PCT/US2014/062913 totreat a disorder, e.g., a disorder described herein. In one embodiment,the ERK1/2 ATP competitive inhibitor is a compound disclosed inInternational Patent Application No. PCT/US2014/062913. In oneembodiment, a TIM-3 antibody molecule is used in combination withCompound A51 or a compound disclosed in International Patent ApplicationNo. PCT/US2014/062913 to treat a disorder such as a cancer.

In some embodiments, the TIM-3 antibody molecule is administerd incombination with one or more agents selected from, Compound A8, CompoundA17, Compound A23, Compound A24, Compound A27, Compound A29, andCompound A33.

In some embodiments, a TIM-3 antibody molecule is administered incombination with an anti-cancer agent having a known activity in animmune cell assay, e.g., in one or more of a huMLR assay, a T cellproliferation assay, and a B-cell proliferation assay. Exemplary assaysare described below. Based on the assay, an IC50 for can be calculatedfor each test agent. In embodiments, the anti-cancer agent has an IC50of, e.g., 0-1 μM, 1-4 μM, or greater than 4 μM, e.g., 4-10 μM or 4-20μM. In embodiments, the second therapeutic agent is chosen from one ormore of: Compound A9, Compound A16, Compound A17, Compound A21, CompoundA22, Compound A25, Compound A28, Compound A48, and Compound 49.

In some embodiments, the Compound A28 (or a compound related to CompoundA28) is administered at a dose of approximately 5-10 or 10-30 mg. Insome embodiments, the Compound A22 (or compound related to Compound A22)is administered at a dose of about 200 mg. In some embodiments, theCompound A17 (or compound related to Compound A17) is administered at adose of approximately 400-600 mg. In some embodiments, the Compound A16(or compound related to Compound A16) is administered at a dose ofapproximately 400-600 mg PO qDay. In some embodiments, the Compound A29(or compound related to Compound A29) is administered at a dose ofapproximately 200-400 or 300-400 mg. In some embodiments, the CompoundA24 (or compound related to Compound A24) is administered at a dose ofapproximately 200-600 mg. In some embodiments, the Compound A23(ceritinib) (or compound related to ceritinib) is administered at a doseof approximately 750 mg once daily. In some embodiments, the Compound A8(or compound related to Compound A8) is administered at a dose ofapproximately 200-400 or 300-400 mg. In some embodiments, the CompoundA5 (or compound related to Compound A5) is administered at a dose ofapproximately 100-125 mg. In some embodiments, the Compound A6 (orcompound related to Compound A6) is administered at a dose of about 100mg. In some embodiments, the Compound A1 (or compound related toCompound A1) is administered at a dose of approximately 200-300 or200-600 mg. In some embodiments, the Compound A40 (or compound relatedto Compound A40) is administered at a dose of approximately 150-250 mg.In embodiments, the Compound A10 (or compound related to Compound A10)is administered at a dose of approximately 400 to 700 mg, e.g.,administered three times weekly, 2 weeks on and one week off. Inembodiments, the BCR-ABL inhibitor is administered at a dose ofapproximately 20 mg bid-80 mg bid.

Exemplary huMLR assay and B or T cell proliferation assays are providedbelow. Human mixed lymphocyte reaction

The Mixed Lymphocyte Reaction (MLR) is a functional assay which measuresthe proliferative response of lymphocytes from one individual (theresponder) to lymphocytes from another individual (the stimulator). Toperform an allogeneic MLR, peripheral blood mononuclear cells (PBMC)from three donors were isolated from buffy-coats of unknown HLA type(Kantonspital Blutspendezentrum from Bern and Aarau, Switzerland). Thecells were prepared at 2.105 in 0.2 mL of culture medium containing RPMI1640 GlutaMAX™ with 10% fetal calf serum (FCS), 100U penicillin/100 μgstreptomycin, 50 μM 2-Mercaptoethanol. Individual 2-way reactions wereset up by mixing PBMC from two different donors at a 1:1 ratio andco-cultures were done in triplicates in flat-bottomed 96-well tissueculture plates for 6 days at 37° C., 5% CO2, in presence or not of an8-point concentration range of test compounds. Cells were pulsed with3H-TdR (1 μCi/0.2 mL) for the last 16 h of culture and incorporatedradioactivity was used as a measure of cell proliferation. Theconcentration that inhibited 50% of the maximal huMLR response (IC50)was calculated for each compound. Cyclosporine was used as a positivecontrol of huMLR inhibition.

Human B Cell Proliferation Assay

PBMC were freshly isolated by Ficoll-Paque density gradient from humanblood and subjected to negative B-cell isolation. B cells wereresuspended in culture medium (RPMI 1640, HEPES, 10% FCS, 50 μg/mLgentamicine, 50 μM 2-Mercaptoethanol, 1× ITS (Insulin, Transferrin andSodium Selenite), 1× Non-Essential Amino-Acids) at a concentration of9.104 per well in a flat-bottom 96-well culture plate. B cellstimulation was performed by human anti-IgM antibody molecule (30 ug/mL)and IL-4 (75 ng/mL) or by CD40 ligand (3 ug/mL) and IL-4 (75 ng/mL) inpresence or not of a 7-point concentration range of test compounds.After 72 h of culture at 37° C., 10% CO2, cells were pulsed with 3H-TdR(1 μCi/well) for the last 6 h of culture. B cells were then harvestedand the incorporation of thymidine was measured using a scintillationcounter. Of each duplicate treatment, the mean was calculated and thesedata were plotted in XLfit 4 to determine the respective IC50 values.

Human T Cell Proliferation Assay

PBMC were freshly isolated by Ficoll-Paque density gradient from humanblood and subjected to negative isolation of T cells. T cells wereprepared in culture medium (RPMI 1640, HEPES, 10% FCS, 50 μg/mLgentamicine, 50 μM 2-Mercaptoethanol, 1× ITS (Insulin, Transferrin andSodium Selenite), 1× Non-Essential Amino-Acids) at a concentration of8.104 per well in a flat-bottom 96-well culture plate. T cellstimulation was performed by human anti-CD3 antibody molecule (10 ug/mL)or by human anti-CD3 antibody molecule (5 μg/mL) and anti-CD28 antibodymolecule (1 μm/mL) in presence or not of a 7-point concentration rangeof test compounds. After 72 h of culture at 37° C., 10% CO2, cells werepulsed with 3H-TdR (1 μCi/well) for the last 6 h of culture. Cellproliferation was measured by the incorporation of thymidine allowingIC50 determination for each tested compound.

Down-Modulators of the Immune System

In an alternative embodiment, the anti-TIM-3 antibody moleculesdisclosed herein are used to produce anti-idiotypic peptides orantibodies (Wallmann, J. et al. (2010) “Anti-Ids in Allergy: Timelinessof a Classic Concept,” World Allergy Organiz. J. 3(6):195-201; Nardi, M.et al. (2000) “Antiidiotype Antibody Against Platelet Anti-GpiiiaContributes To The Regulation Of Thrombocytopenia In HIV-1-ITPPatients,” J. Exp. Med. 191(12):2093-2100) or mimetics (Zang, Y. C. etal. (2003) “Human Anti-Idiotypic T Cells Induced By TCR PeptidesCorresponding To A Common CDR3Sequence Motif In Myelin BasicProtein-Reactive T Cells,” Int. Immunol. 15(9):1073-1080; Loiarro, M. etal. (Epub 2010 Apr. 8) “Targeting TLR/IL-1R Signalling In HumanDiseases,” Mediators Inflamm. 2010:674363) of TIM-3. Such moleculesserve as surrogates for TIM-3, and thus their administration to asubject down-modulates the immune system of such subject by mimicking orfacilitating ligand-TIM-3 binding. Such molecules have utility in thetreatment of graft vs. host disease. Similarly, agonist antibodies thati) enhance binding between such antibodies and such receptor/ligand orii) trigger signal transduction when bound directly to a TIM-3 ligand orTIM-3, have utility as agonists of TIM-3 signaling and thus have utilityin the treatment of inflammation and autoimmune disease, by directly orindirectly agonizing receptor activity.

Bispecific antibodies, exhibiting immunospecific binding to both TIM-3and TIM-3 ligands are capable of binding to both APC and T-cells, andthus facilitate the co-localization of APCs and T-cells. Suchco-localization facilitates the ability of such cells to bind togethervia TIM-3 ligand and TIM-3 molecules that are not complexed withantibody, or by co-inhibitory molecules. Such binding provides downmodulation of the immune system of the recipient.

Down-modulation of the immune system is desirable in the treatment ofinflammatory and auto-immune diseases, and graft vs. host disease(GvHD). Examples of autoimmune disorders that may be treated byadministering the antibodies of the present invention include, but arenot limited to, alopecia greata, ankylosing spondylitis,antiphospholipid syndrome, autoimmune Addison's disease, autoimmunediseases of the adrenal gland, autoimmune hemolytic anemia, autoimmunehepatitis, autoimmune oophoritis and orchitis, autoimmunethrombocytopenia, Behcet's disease, bullous pemphigoid, cardiomyopathy,celiac sprue-dermatitis, chronic fatigue immune dysfunction syndrome(CFIDS), chronic inflammatory demyelinating polyneuropathy,Churg-Strauss syndrome, cicatrical pemphigoid, CREST syndrome, coldagglutinin disease, Crohn's disease, discoid lupus, essential mixedcryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis,Graves' disease, Guillain-Barre, Hashimoto's thyroiditis, idiopathicpulmonary fibrosis, idiopathic thrombocytopenia purpura (ITP), IgAneuropathy, juvenile arthritis, lichen planus, lupus erthematosus,Meniere's disease, mixed connective tissue disease, multiple sclerosis,Neuromyelitis optica (NMO), type 1 or immune-mediated diabetes mellitus,myasthenia gravis, pemphigus vulgaris, pernicious anemia, polyarteritisnodosa, polychrondritis, polyglandular syndromes, polymyalgiarheumatica, polymyositis and dermatomyositis, primaryagammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriaticarthritis, Raynauld's phenomenon, Reiter's syndrome, Rheumatoidarthritis, sarcoidosis, scleroderma, Sjogren's syndrome, stiff-mansyndrome, systemic lupus erythematosus, lupus erythematosus, takayasuarteritis, temporal arteristis/giant cell arteritis, transversemyelitis, ulcerative colitis, uveitis, vasculitides such as dermatitisherpetiformis vasculitis, vitiligo, and Wegener's granulomatosis.

Examples of inflammatory disorders which can be prevented, treated ormanaged in accordance with the methods of the invention include, but arenot limited to, asthma, encephilitis, inflammatory bowel disease,chronic obstructive pulmonary disease (COPD), allergic disorders, septicshock, pulmonary fibrosis, undifferentiated spondyloarthropathy,undifferentiated arthropathy, arthritis, inflammatory osteolysis, andchronic inflammation resulting from chronic viral or bacterialinfections.

Thus, the antibodies and antigen-binding fragments of the presentinvention have utility in the treatment of inflammatory and autoimmunediseases.

Diagnostic Uses

In some aspects, the present disclosure provides a diagnostic method fordetecting the presence of a TIM-3 protein in vitro (e.g., in abiological sample, such as a tissue biopsy, e.g., from a canceroustissue) or in vivo (e.g., in vivo imaging in a subject). The methodincludes: (i) contacting the sample with an antibody molecule describedherein, or administering to the subject, the antibody molecule;(optionally) (ii) contacting a reference sample, e.g., a control sample(e.g., a control biological sample, such as plasma, tissue, biopsy) or acontrol subject with an antibody molecule described herein; and (iii)detecting formation of a complex between the antibody molecule, and thesample or subject, or the control sample or subject, wherein a change,e.g., a statistically significant change, in the formation of thecomplex in the sample or subject relative to the control sample orsubject is indicative of the presence of TIM-3 in the sample. Theantibody molecule can be directly or indirectly labeled with adetectable substance to facilitate detection of the bound or unboundantibody. Suitable detectable substances include various enzymes,prosthetic groups, fluorescent materials, luminescent materials andradioactive materials, as described above and described in more detailbelow.

The term “sample,” as it refers to samples used for detectingpolypeptides includes, but is not limited to, cells, cell lysates,proteins or membrane extracts of cells, body fluids such as blood, ortissue samples.

Complex formation between the antibody molecule and TIM-3 can bedetected by measuring or visualizing either the antibody molecule boundto the TIM-3 antigen or unbound antibody molecule. Any suitabledetection assays can be used, and conventional detection assays includean enzyme-linked immunosorbent assays (ELISA), a radioimmunoassay (RIA)or tissue immunohistochemistry. Alternative to labeling the antibodymolecule, the presence of TIM-3 can be assayed in a sample by acompetition immunoassay utilizing standards labeled with a detectablesubstance and an unlabeled antibody molecule. In this assay, thebiological sample, the labeled standards and the antibody molecule arecombined and the amount of labeled standard bound to the unlabeledbinding molecule is determined. The amount of TIM-3 in the sample isinversely proportional to the amount of labeled standard bound to theantibody molecule.

In some aspects, the present disclosure provides methods of using ananti-TIM-3 antibody molecule to diagnose sepsis, SIRS (SystemicInflammatory Response Syndrome), preeclampsia, or glomerulonephritis.Sepsis is often accompanied by a downregulation of TIM-3 (Yang et al., JImmunol. 2013 Mar. 1; 190(5):2068-79) so lowered levels of TIM-3 areindicative of sepsis while normal levels of TIM-3 are an indication thatsepsis is not present. In SIRS and preeclampsia, TIM-3 levels aredownregulated in peripheral lymphocytes (Miko et al., PLoS ONE 8(8):e71811), so lowered levels of TIM-3 are indicative of SIRS orpreeclampsia, while normal levels of TIM-3 are an indication that SIRSand preeclampsia are not present. In glomerulonephritis, TIM-3 can beupregulated (see Schroll et al., Am J Pathol 2010 April;176(4):1716-1742) so elevated levels of TIM-3 are indicative ofglomerulonephritis, while normal levels are an indication thatglomerulonephritis is not present.

Nucleic Acids

The present disclosure also features nucleic acids comprising nucleotidesequences that encode heavy and light chain variable regions and CDRs ofthe anti-TIM-3 antibody molecules, as described herein. For example, thepresent disclosure features a first and second nucleic acid encodingheavy and light chain variable regions, respectively, of an anti-TIM-3antibody molecule chosen from one or more of the antibody moleculesdisclosed herein, e.g., an antibody of Tables 1-4. The nucleic acid cancomprise a nucleotide sequence encoding any one of the amino acidsequences in the tables herein, or a sequence substantially identicalthereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or moreidentical thereto, or which differs by no more than 3, 6, 15, 30, or 45nucleotides from the sequences provided in Tables 1-4.

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs from a heavy chainvariable region having an amino acid sequence as set forth in Tables1-4, or a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or havingone or more substitutions, e.g., conserved substitutions). In someembodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, or three CDRs from a light chain variableregion having an amino acid sequence as set forth in Tables 1-4, or asequence substantially homologous thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, and/or having one ormore substitutions, e.g., conserved substitutions). In some embodiments,the nucleic acid can comprise a nucleotide sequence encoding at leastone, two, three, four, five, or six CDRs from heavy and light chainvariable regions having an amino acid sequence as set forth in Tables1-4, or a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or havingone or more substitutions, e.g., conserved substitutions).

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs from a heavy chainvariable region having the nucleotide sequence as set forth in Tables1-4, a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or capableof hybridizing under the stringency conditions described herein). Insome embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, or three CDRs from a light chain variableregion having the nucleotide sequence as set forth in Tables 1-4, or asequence substantially homologous thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, and/or capable ofhybridizing under the stringency conditions described herein). Incertain embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, three, four, five, or six CDRs from heavyand light chain variable regions having the nucleotide sequence as setforth in Tables 1-4, or a sequence substantially homologous thereto(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, and/or capable of hybridizing under the stringency conditionsdescribed herein). The nucleic acids disclosed herein includedeoxyribonucleotides or ribonucleotides, or analogs thereof. Thepolynucleotide may be either single-stranded or double-stranded, and ifsingle-stranded may be the coding strand or non-coding (antisense)strand. A polynucleotide may comprise modified nucleotides, such asmethylated nucleotides and nucleotide analogs. The sequence ofnucleotides may be interrupted by non-nucleotide components. Apolynucleotide may be further modified after polymerization, such as byconjugation with a labeling component. The nucleic acid may be arecombinant polynucleotide, or a polynucleotide of genomic, cDNA,semisynthetic, or synthetic origin which either does not occur in natureor is linked to another polynucleotide in a nonnatural arrangement.

In some aspects, the application features host cells and vectorscontaining the nucleic acids described herein. The nucleic acids may bepresent in a single vector or separate vectors present in the same hostcell or separate host cell, as described in more detail hereinbelow.

Vectors

Further provided herein are vectors comprising nucleotide sequencesencoding an antibody molecule described herein. In some embodiments, thevectors comprise nucleotides encoding an antibody molecule describedherein. In some embodiments, the vectors comprise the nucleotidesequences described herein. The vectors include, but are not limited to,a virus, plasmid, cosmid, lambda phage or a yeast artificial chromosome(YAC).

Numerous vector systems can be employed. For example, one class ofvectors utilizes DNA elements which are derived from animal viruses suchas, for example, bovine papilloma virus, polyoma virus, adenovirus,vaccinia virus, baculovirus, retroviruses (Rous Sarcoma Virus, MMTV orMOMLV) or SV40 virus. Another class of vectors utilizes RNA elementsderived from RNA viruses such as Semliki Forest virus, Eastern EquineEncephalitis virus and Flaviviruses.

Additionally, cells which have stably integrated the DNA into theirchromosomes may be selected by introducing one or more markers whichallow for the selection of transfected host cells. The marker mayprovide, for example, prototropy to an auxotrophic host, biocideresistance, (e.g., antibiotics), or resistance to heavy metals such ascopper, or the like. The selectable marker gene can be either directlylinked to the DNA sequences to be expressed, or introduced into the samecell by cotransformation. Additional elements may also be needed foroptimal synthesis of mRNA. These elements may include splice signals, aswell as transcriptional promoters, enhancers, and termination signals.

Once the expression vector or DNA sequence containing the constructs hasbeen prepared for expression, the expression vectors may be transfectedor introduced into an appropriate host cell. Various techniques may beemployed to achieve this, such as, for example, protoplast fusion,calcium phosphate precipitation, electroporation, retroviraltransduction, viral transfection, gene gun, lipid based transfection orother conventional techniques. In the case of protoplast fusion, thecells are grown in media and screened for the appropriate activity.

Methods and conditions for culturing the resulting transfected cells andfor recovering the antibody molecule produced are known to those skilledin the art, and may be varied or optimized depending upon the specificexpression vector and mammalian host cell employed, based upon thepresent description.

Cells

The present disclosure also provides host cells comprising a nucleicacid encoding an antibody molecule as described herein.

In some embodiments, the host cells are genetically engineered tocomprise nucleic acids encoding the antibody molecule.

In certain embodiments, the host cells are genetically engineered byusing an expression cassette. The phrase “expression cassette,” refersto nucleotide sequences, which are capable of affecting expression of agene in hosts compatible with such sequences. Such cassettes may includea promoter, an open reading frame with or without introns, and atermination signal. Additional factors necessary or helpful in effectingexpression may also be used, such as, for example, an induciblepromoter.

The disclosure also provides host cells comprising the vectors describedherein.

The cell can be, but is not limited to, a eukaryotic c

ell, a bacterial cell, an insect cell, or a human cell. Suitableeukaryotic cells include, but are not limited to, Vero cells, HeLacells, COS cells, CHO cells, HEK293 cells, BHK cells and MDCKII cells.Suitable insect cells include, but are not limited to, Sf9 cells.

Exemplary sequences of anti-TIM-3 antibodies are described in the Tables1-4 below.

TABLE 1 Summary of the sequences of the murine antibody ABTIM3. Antibodydesignation SEQ ID NO Description ABTIM3 1 VH amino acid sequence 2 VLamino acid sequence 3 VHCDR1 amino acid sequence 4 VHCDR2 amino acidsequence 5 VHCDR3 amino acid sequence 6 VLCDR1 amino acid sequence 7VLCDR2 amino acid sequence 8 VLCDR3 amino acid sequence

TABLE 2  Depiction of the amino acid sequences of themurine antibody ABTIM3 heavy chain variable  domain and light chain variable domain. SEQ ID NO Sequence 1QVQLQQPGAE LVKPGASVKM SCKASGYTFT SYNMHWIKQTPGQGLEWIGD IYPGNGDTSY NQKFKGKATL TADKSSSTVY MQLSSLTSED SAVYYCARVG GAFPMDYWGQ GTSVTVSS 2DIVLTQSPAS LAVSLGQRAT ISCRASESVE YYGTSLMQWYQQKPGQPPKL LIYAASNVES GVPARFSGSG SGTDFSLNIH PVEEDDIAIY FCQQSRKDPS TFGGGTKLEI K CDRs are shown in white text on ablack background.

TABLE 3  Depiction of the amino acid sequences of themurine antibody ABTIM3 heavy chain CDRs and light chain CDRs. SEQ ID NOSequence 3 SYNMH 4 DIYPGNGDTSYNQKFKG 5 VGGAFPMDY 6 RASESVEYYGTSLMQ 7AASNVES 8 QQSRKDPST

Exemplary sequences of anti-TIM-3 antibodies are described in Table 4.The antibody molecules include murine ABTIM3, and humanized antibodymolecules. The amino acid and nucleotide sequences of the heavy andlight chain CDRs, the heavy and light chain variable regions, and theheavy and light chains are shown.

TABLE 4  Summary of the sequences of exemplary anti-TIM-3 antibodies.Hybridoma clone ABTIM3 SEQ ID NO: 3 HCDR1 SYNMH (Kabat) SEQ ID NO: 4HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Kabat)SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2 YPGNGD 10(Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: 1 VHQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWIKQTPGQGLEWIGDIYPGNGDTSYNQKFKGKATLTADKSSSTVYMQLSSLTSEDSAVYYCARVGGAFP MDYWGQGTSVTVSSSEQ ID NO: 11 DNA VHCAGGTGCAACTGCAGCAGCCTGGGGCTGAGCTGGTGAAGCCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACATTTACCAGTTACAATATGCACTGGATAAAGCAGACACCTGGACAGGGCCTGGAATGGATTGGAGATATTTATCCAGGAAATGGTGATACTTCCTACAATCAGAAATTCAAAGGCAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGTCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGAGTGGGGGGTGCCTTTCCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA SEQ ID NO: 6 LCDR1RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8LCDR3 QQSRKDPST (Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia)SEQ ID NO: LCDR2 AAS 13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia)SEQ ID NO: 2 VL DIVLTQSPASLAVSLGQRATISCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPARFSGSGSGTDFSLNIHPVEEDDIAIYFCQQSRKDPSTFGG GTKLEIKSEQ ID NO: 15 DNA VLGACATTGTGCTCACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGAGCCACCATCTCCTGCAGAGCCAGTGAAAGTGTTGAATATTATGGCACAAGTTTAATGCAGTGGTACCAACAGAAACCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACGTAGAATCTGGGGTCCCTGCCAGGTTTAGTGGCAGTGGGTCTGGGACAGACTTCAGCCTCAACATCCATCCTGTGGAGGAGGATGATATTGCAATATATTTCTGTCAGCAAAGTAGGAAGGATCCTTCGACGTTCGGTGGAGGCACCAAGCTGGAGATCAAA ABTIM3-hum01 SEQ ID NO: 3 HCDR1 SYNMH (Kabat)SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 16PGNGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG17 TGAAAGTCTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCAGGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 18 ChainPGNGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG 19 HeavyTGAAAGTCTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTTCGCCAGGCCCCAGGGCAGGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 20YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 21GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 22 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 23Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC AETIM3-hum02 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGSGD 25 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 26PGSGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG27 TGAAAGTTAGCTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAAGATACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 28 ChainPGSGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG 29 HeavyTGAAAGTTAGCTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTTCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAAGATACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 20YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 21GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 22 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 23Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC AETIM3-hum03 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 32PGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG33 TGAAAGTTAGCTGTAAAGCTAGTGGCTATACTTTCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 34 ChainPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG 35 HeavyTGAAAGTTAGCTGTAAAGCTAGTGGCTATACTTTCACTTCTTATAATATGCA ChainCTGGGTCCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 20YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 21GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 22 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 23Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum04 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) (Chothia) SEQ ID NO: HCDR2 YPGNGD 10 (Chothia)SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 36PGNGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG37 TGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCAGGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGGCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 38 ChainPGNGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG 39 HeavyTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGATTAGACAGGCCCCAGGGCAGGGCCTCGAGTGGATCGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGGCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 40YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 41GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 42 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 43Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum05 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGSGD 25(Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 44PGSGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCAAGCG45  TTAAAGTCTCATGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGGCAAGGCCTGGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGAGTGAAGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCAAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 46 ChainPGSGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCAAGCG 47 HeavyTTAAAGTCTCATGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGATTAGACAGGCCCCAGGGCAAGGCCTGGAGTGGATCGGCGATATCTACCCCGGTAGCGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGAGTGAAGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCAAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 40YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 41GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 42 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 43Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum06 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 48PGQGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG49 TGAAAGTCTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGATTAGACAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 50 ChainPGQGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTG 51 HeavyTGAAAGTCTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGATTAGACAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGCTACCCTGACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 40YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 41GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 42 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGC 43Light GGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum07 (Kabat) SEQ ID NO: 4 HCDR2DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Kabat)SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2 YPGNGD10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 36PGNGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTCCAGCTGGTCCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG115 TGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCACTGGATCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGAAACGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCCTGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAGCTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCTCCTCC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWIRQAPGQGLEWIGDIY 116 ChainPGNGDTSYNQKFKGRATLTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTCCAGCTGGTCCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG 117 HeavyTGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCA ChainCTGGATCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGAAACGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCCTGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAGCTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCTCCTCCGCTAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCATGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 20YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA 118GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAGTCTGATGCAGTGGTACCAGCAGAAGCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 22 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA 119Light GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAG ChainTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAGCGGACTGTTGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGTTCACCGGTGACAAAGAGCTTCAACAGGGGAGAGTGT ABTIM3-hum08 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 16PGNGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTCCAGCTGGTCCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG120 TGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCACTGGGTCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGAAACGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAGCTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCTCCTCC SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 121 ChainPGNGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTCCAGCTGGTCCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG 122 HeavyTGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCA ChainCTGGGTCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGAAACGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAGCTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCTCCTCCGCTAGCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCATGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 40 GTKVEIKSEQ ID NO: DNA VL GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA123 GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAGTCTGATGCAGTGGTACCAGCAGAAGCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTTTTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 42 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA 124Light GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAG ChainTCTGATGCAGTGGTACCAGCAGAAGCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTTTTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAGCGGACTGTTGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGTTCACCGGTGACAAAGAGCTTCAACAGGGGAGAGTGT LABTIM3-hum09 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 52PGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCG53 TGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 54 ChainPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCG 55 HeavyTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGA 57GAGCTACACTGAGCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGGATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGA 59Light GAGCTACACTGAGCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGGATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum10 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 60PGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCAC61 TGAAGATTAGCTGTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCCTGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 62 ChainPGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNAGAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCAC 63 HeavyTGAAGATTAGCTGTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTCCGCCAGATGCCCGGGAAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCCTGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATCGTCCTGACTCAGTCACCCCGTACCCTGAGCCCTGGCGAGA 57GAGCTACACTGAGCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGGATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATCGTCCTGACTCAGTCACCCGCTACCCTGAGCCTGAGCCCTGGCGAGA 59Light GAGCTACACTGAGCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAAGCCCCTAGACTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGGATCCCCGCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGGAACCCGAGGATATCGCCGTCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum11 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 52 PGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCG53 TGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 54 ChainPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCG 55 HeavyTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATA 65GAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATA 67Light GAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum12 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 60PGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCAC61 TGAAGATTAGCTGTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCACTGGGTCCGCCAGATGCCCGGGAAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCCTGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGC SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 62 ChainPGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: DNAGAAGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAGCCCGGCGAGTCAC 63 HeavyTGAAGATTAGCTGTAAAGGTTCAGGCTACACCTTCACTAGCTATAATATGCA ChainCTGGGTCCGCCAGATGCCCGGGAAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGGCAAGTCACAATTAGCGCCGATAAGTCTATTAGCACCGTCTACCTGCAGTGGTCTAGCCTGAAGGCTAGTGACACCGCTATGTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATA 65GAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATA 67Light GAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum13 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGSGD 25 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 68PGSGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG69 TCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 70 ChainPGSGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG 71 HeavyTCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCA ChainCTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 125GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAA SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 126Light GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAG ChainCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT AETIM3-hum14 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 72PGQGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG73 TCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 74 ChainPGQGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG 75 HeavyTCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCA ChainCTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDRI SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64 YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 125GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAA SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 126Light GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAG ChainCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum15 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Kabat) (Chothia) SEQ ID NO: HCDR2 YPGSGD 25 (Chothia)SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 76PGSGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT77 TGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 78 ChainPGSGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNAGAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT 79 HeavyTGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCA ChainCTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 127GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 128Light GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATC ChainCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum16 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 80PGQGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT81 TGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 82 ChainPGQGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNAGAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT 83 HeavyTGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCA ChainCTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 127GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 128Light GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATC ChainCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum17 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGSGD 25 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGSGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP 68 MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG69 TCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 70 ChainPGSGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG 71 HeavyTCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCA ChainCTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCAGTGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 127GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 128Light GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATC ChainCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum18 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 72PGQGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG73 TCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCACTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 74 ChainPGQGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNACAGGTGCAATTGGTTCAGTCAGGAGCAGAAGTTAAGAAGCCAGGATCATCCG 75 HeavyTCAAGGTGTCCTGCAAAGCATCTGGCTACACCTTCACCAGCTACAATATGCA ChainCTGGGTCCGACAAGCCCCTGGGCAGGGCTTGGAGTGGATGGGAGACATTTACCCCGGCCAGGGTGACACTTCCTATAACCAGAAGTTCAAGGGCCGAGTCACTATTACCGCTGACAAGTCCACCTCCACAGTCTACATGGAACTCTCTTCTCTGAGATCCGAGGACACTGCCGTCTATTACTGCGCTCGCGTGGGCGGTGCTTTCCCAATGGACTATTGGGGACAGGGCACAACCGTGACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 56YAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 127GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATCCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAG SEQ ID NO: LightEIVLTQSPATLSLSPGERATLSCRASESVEYYGTSLMQWYQQKPGQAPRLLI 58 ChainYAASNVESGIPARFSGSGSGTDFTLTISSLEPEDIAVYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GAGATTGTTCTTACGCAAAGTCCCGCCACACTTAGTTTGTCACCAGGAGAGC 128Light GCGCCACCCTGAGCTGCAGAGCTTCAGAGAGTGTGGAATACTACGGCACATC ChainCCTGATGCAGTGGTATCAGCAGAAACCAGGACAGGCTCCTCGGCTGCTGATCTACGCAGCCAGCAACGTCGAGTCCGGCATTCCAGCCAGATTTTCTGGGTCAGGATCTGGAACTGACTTTACACTGACAATCTCCAGCCTGGAACCCGAGGACATTGCTGTGTATTTTTGTCAACAGTCCCGGAAGGACCCCAGTACCTTTGGAGGTGGAACCAAGGTAGAGATAAAGCGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum19 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGSGDTSYNQKFKG 24 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGSGD 25 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 76PGSGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT77 TGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 78 ChainPGSGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNAGAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT 79 HeavyTGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCA ChainCTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAGTGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 125GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAA SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 126Light GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAG ChainCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum20 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 80PGQGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH GAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT81 TGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: HeavyEVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 82 ChainPGQGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK SEQ ID NO: DNAGAAGTTCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT 83 HeavyTGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCA ChainCTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCCAGGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCCTCTACAAAGGGCCCCTCCGTCTTTCCACTCGCGCCGTGCTCTCGCTCCACCTCAGAGTCAACTGCCGCTCTGGGTTGCCTGGTCAAGGACTACTTCCCAGAGCCGGTGACAGTGAGCTGGAACAGTGGGGCCCTGACATCCGGCGTTCATACCTTCCCCGCAGTCCTCCAGTCCTCAGGCCTGTATTCCCTGAGCAGCGTTGTCACAGTGCCCTCCAGCTCTCTTGGCACGAAAACCTACACATGCAACGTTGATCATAAGCCGTCTAATACCAAGGTGGATAAAAGAGTGGAGAGCAAGTACGGCCCACCCTGCCCGCCTTGCCCAGCTCCGGAGTTCCTGGGCGGACCATCCGTTTTCTTGTTTCCACCCAAACCTAAAGACACTCTGATGATTTCCCGAACCCCTGAAGTGACTTGCGTTGTGGTGGACGTCTCCCAGGAGGACCCAGAAGTGCAATTCAACTGGTACGTGGACGGGGTGGAGGTGCACAATGCAAAAACCAAACCAAGGGAGGAACAGTTTAATTCAACATATAGGGTTGTGTCTGTGCTGACGGTTCTGCATCAGGACTGGCTGAACGGAAAGGAATACAAGTGCAAGGTGTCCAACAAAGGACTGCCAAGCTCTATCGAGAAAACAATCTCTAAGGCCAAGGGACAACCTAGAGAGCCCCAAGTTTACACCCTGCCACCATCACAGGAAGAGATGACCAAAAATCAGGTGAGCTTGACATGCCTGGTGAAGGGCTTCTACCCTAGCGATATTGCGGTTGAGTGGGAGTCAAATGGCCAGCCTGAGAACAACTATAAGACTACTCCTCCCGTGCTGGACTCCGACGGGAGCTTTTTCCTGTATTCCAGGCTTACAGTCGATAAGAGCAGATGGCAAGAGGGGAATGTGTTTTCCTGCTCCGTGATGCACGAGGCTCTCCATAACCATTATACTCAGAAAAGTCTCTCTCTGTCACTGGGCAAA SEQ ID NO: 6 LCDR1 RASESVEYYGTSLMQ(Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8 LCDR3 QQSRKDPST(Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia) SEQ ID NO: LCDR2 AAS13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia) SEQ ID NO: VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 64YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 125GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAA SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 66 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 126Light GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAG ChainCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAACGTACGGTGGCAGCTCCGTCTGTTTTCATCTTTCCACCTAGCGACGAGCAACTCAAAAGTGGTACAGCATCCGTGGTTTGTCTGCTGAACAATTTTTACCCCAGGGAGGCTAAGGTCCAGTGGAAAGTCGATAACGCTCTTCAGTCTGGCAACAGTCAGGAGAGCGTCACAGAGCAGGACTCTAAGGATAGCACTTATAGTCTGTCCTCCACGCTGACACTGTCTAAAGCGGATTATGAGAAGCACAAGGTTTACGCCTGTGAGGTAACGCACCAAGGACTCTCCTCCCCAGTTACCAAATCTTTCAACAGAGGAGAATGT ABTIM3-hum21 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 84PGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTLVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTGCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG85 TGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCACTGGGTCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGACAGGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAACTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIY 86 ChainPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: DNACAGGTGCAATTGGTGCAGAGCGGAGCAGAGGTCAAAAAGCCCGGAGCAAGCG 87 HeavyTGAAGGTCTCATGCAAAGCAAGCGGATACACATTTACATCATACAACATGCA ChainCTGGGTCAGGCAGGCTCCAGGACAGGGACTGGAGTGGATCGGGGACATCTACCCTGGACAGGGCGATACTAGCTATAATCAGAAGTTCAAAGGCCGGGCCACCATGACAGCTGACAAGTCTACTAGTACCGTGTATATGGAACTGAGCTCCCTGCGGTCTGAAGATACCGCAGTGTACTATTGCGCCAGAGTCGGGGGGGCATTTCCTATGGATTATTGGGGGCAGGGGACTCTGGTCACTGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCTGCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 LCDR1RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8LCDR3 QQSRKDPST (Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia)SEQ ID NO: LCDR2 AAS 13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia)SEQ ID NO: VL DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 88YAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA 89GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAGTCTGATGCAGTGGTACCAGCAGAAGCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAG SEQ ID NO: LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLI 90 ChainYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GACATCGTCCTGACACAGTCTCCTGACAGCCTGGCAGTGAGCCTGGGCGAAA 91Light GGGCAACCATTAATTGTAGAGCTTCCGAGTCCGTCGAGTACTATGGCACTAG ChainTCTGATGCAGTGGTACCAGCAGAAGCCAGGGCAGCCCCCTAAACTGCTGATCTATGCAGCTAGCAACGTGGAGTCCGGAGTCCCAGACCGGTTCTCTGGAAGTGGGTCAGGAACCGATTTTACCCTGACAATTAGCTCCCTGCAGGCAGAAGACGTGGCCGTCTACTATTGTCAGCAGAGCCGCAAGGACCCAAGCACATTCGGAGGGGGGACCAAAGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum22 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: HCDR2 DIYPGQGDTSYNQKFKG 30 (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGQGD 31 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 92PGQGDTSYNQKFKGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCAGGCGCCAGCG93 TGAAGGTGTCCTGCAAGGCCAGCGGCTACACCTTTACCAGCTACAACATGCACTGGGTGCGCCAGGCCCCTGGACAGGGACTGGAATGGATGGGCGACATCTACCCCGGCCAGGGCGACACCTCCTACAACCAGAAATTCAAGGGCAGAGTGACCATGACCCGGGACACCAGCACCTCCACCGTGTACATGGAACTGAGCAGCCTGCGGAGCGAGGACACCGCCGTGTACTACTGTGCTAGAGTGGGCGGAGCCTTCCCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIY 94 ChainMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: DNACAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCAGGCGCCAGCG 95 HeavyCTGGGTGCGCCAGGCCCCTGGACAGGGACTGGAATGGATGGGCGACATCTAC ChainCCCGGCCAGGGCGACACCTCCTACAACCAGAAATTCAAGGGCAGAGTGACCATGACCCGGGACACCAGCACCTCCACCGTGTACATGGAACTGAGCAGCCTGCGGAGCGAGGACACCGCCGTGTACTACTGTGCTAGAGTGGGCGGAGCCTTCCCCATGGACTATTGGGGCCAGGGCACCACCGTGACCGTGAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCTGCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 LCDR1RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8LCDR3 QQSRKDPST (Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia)SEQ ID NO: LCDR2 AAS 13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia)SEQ ID NO: VL AIRLTQSPSSFSASTGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 96YAASNVESGVPSRFSGSGSGTDFTLTISSLQSEDFATYYCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCCATCAGACTGACCCAGAGCCCCAGCTCCTTTAGCGCCAGCACCGGCGACA 97GAGTGACCATCACCTGTAGAGCCAGCGAGAGCGTGGAATATTACGGCACCAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGCCGCCAGCAATGTGGAAAGCGGCGTGCCCAGCAGATTCAGCGGCTCTGGCAGCGGCACCGACTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGGACTTCGCCACCTACTACTGCCAGCAGAGCCGGAAGGACCCCAGCACATTTGGCGGAGGCACCAAGGTGGAAATCAAG SEQ ID NO: LightAIRLTQSPSSFSASTGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 98 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQSEDFATYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCCATCAGACTGACCCAGAGCCCCAGCTCCTTTAGCGCCAGCACCGGCGACA 99Light GAGTGACCATCACCTGTAGAGCCAGCGAGAGCGTGGAATATTACGGCACCAG ChainCCTGATGCAGTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGCCGCCAGCAATGTGGAAAGCGGCGTGCCCAGCAGATTCAGCGGCTCTGGCAGCGGCACCGACTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGGACTTCGCCACCTACTACTGCCAGCAGAGCCGGAAGGACCCCAGCACATTTGGCGGAGGCACCAAGGTGGAAATCAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC ABTIM3-hum23 SEQ ID NO: 3 HCDR1 SYNMH(Kabat) SEQ ID NO: 4 HCDR2 DIYPGNGDTSYNQKFKG (Kabat) SEQ ID NO: 5 HCDR3VGGAFPMDY (Kabat) SEQ ID NO: 9 HCDR1 GYTFTSY (Chothia) SEQ ID NO: HCDR2YPGNGD 10 (Chothia) SEQ ID NO: 5 HCDR3 VGGAFPMDY (Chothia) SEQ ID NO: VHQVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 100PGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFP MDYWGQGTTVTVSSSEQ ID NO: DNA VH CAGGTGCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT101 TGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCACTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAATGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCA SEQ ID NO: HeavyQVQLVQSGAEVKKPGESLKISCKGSGYTFTSYNMHWVRQMPGKGLEWMGDIY 102 ChainPGNGDTSYNQKFKGQVTISADKSISTVYLQWSSLKASDTAMYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVAVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: DNACAGGTGCAATTGGTACAGTCTGGCGCAGAAGTAAAGAAACCAGGAGAGAGTT 103 HeavyTGAAAATTTCCTGCAAGGGCAGTGGGTACACATTCACGTCCTACAATATGCA ChainCTGGGTGAGACAGATGCCAGGCAAGGGCCTGGAGTGGATGGGAGACATATACCCAGGCAATGGAGACACAAGCTATAATCAGAAATTCAAAGGACAGGTGACGATCTCCGCAGACAAATCCATATCTACGGTCTACCTCCAGTGGTCCTCACTTAAAGCCTCCGACACCGCCATGTACTATTGCGCTCGGGTAGGTGGCGCGTTTCCAATGGACTATTGGGGCCAAGGGACCACAGTAACCGTCAGCTCAGCTAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGTCCAGCGTGGTGACAGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTGGAGCCCAAGAGCTGCGACAAGACCCACACCTGCCCCCCCTGCCCAGCCCCAGAGCTGCTGGGCGGACCCTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGGCCGTGAGCCACGAGGACCCAGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCCAGAGAGGAGCAGTACAACAGCACCTACAGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAATACAAGTGCAAGGTCTCCAACAAGGCCCTGGCAGCCCCCATCGAAAAGACCATCAGCAAGGCCAAGGGCCAGCCACGGGAGCCCCAGGTGTACACCCTGCCCCCCTCCCGGGAGGAGATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCAGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGAGCCTGAGCCTGTCCCCCGGCAAG SEQ ID NO: 6 LCDR1RASESVEYYGTSLMQ (Kabat) SEQ ID NO: 7 LCDR2 AASNVES (Kabat) SEQ ID NO: 8LCDR3 QQSRKDPST (Kabat) SEQ ID NO: LCDR1 SESVEYYGTSL 12 (Chothia)SEQ ID NO: LCDR2 AAS 13 (Chothia) SEQ ID NO: LCDR3 SRKDPS 14 (Chothia)SEQ ID NO: VL AIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 104YAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGG GTKVEIK SEQ ID NO:DNA VL GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 105GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAGCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAA SEQ ID NO: LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLI 106 ChainYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECSEQ ID NO: DNA GCAATACAGTTGACACAGAGTCCTTCAAGTTTGTCCGCTTCCGTTGGCGACC 107Light GAGTGACAATCACCTGTAGAGCATCCGAGTCAGTGGAGTATTATGGCACTAG ChainCCTGATGCAGTGGTATCAGCAAAAGCCAGGGAAAGCCCCAAAGCTGCTGATATATGCCGCGAGTAACGTCGAGTCAGGGGTGCCATCAAGATTCTCCGGTTCCGGGTCCGGAACCGACTTCACACTGACCATCTCTTCCCTTCAGCCAGAGGACTTCGCTACGTACTTTTGCCAGCAGTCACGGAAAGATCCCTCTACTTTCGGAGGTGGGACAAAAGTCGAAATTAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC

TABLE 5Constant region amino acid sequences of human IgG heavy chains and human kappalight chain HCIgG4 (S228P) mutant constant region amino acid sequence (EU Numbering)ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSVFLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTYRVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTKNQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEGNVFSCSVMHE ALHNHYTQKS LSLSLGK (SEQ ID NO: 108) LCHuman kappa constant region amino acid sequenceRTVAAPSVFI FPPSDEQLKS GTASVVCLLN NFYPREAKVQ WKVDNALQSG NSQESVTEQDSKDSTYSLSS TLTLSKADYE KHKVYACEVT HQGLSSPVTK SFNRGEC (SEQ ID NO: 109) HCIgG4 (S228P) mutant constant region amino acid sequence lacing C-terminallysine (K)(EU Numbering)ASTKGPSVFP LAPCSRSTSE STAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTKT YTCNVDHKPS NTKVDKRVES KYGPPCPPCP APEFLGGPSVFLFPPKPKDT LMISRTPEVT CVVVDVSQED PEVQFNWYVD GVEVHNAKTK PREEQFNSTYRVVSVLTVLH QDWLNGKEYK CKVSNKGLPS SIEKTISKAK GQPREPQVYT LPPSQEEMTKNQVSLTCLVK GFYPSDIAVE WESNGQPENN YKTTPPVLDS DGSFFLYSRL TVDKSRWQEGNVFSCSVMHE ALHNHYTQKS LSLSLG (SEQ ID NO: 110) HC IgG1 wild typeASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGGPSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYNSTYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREEMTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRWQQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 111) HCIgG1 (N297A) mutant constant region amino acid sequence (EU Numbering)ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGGPSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYASTYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREEMTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRWQQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 112) HCIgG1 (D265A, P329A) mutant constant region amino acid sequence (EU Numbering)ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPELLGGPSVFLFPPKP KDTLMISRTP EVTCVVVAVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYNSTYRVVSVLT VLHQDWLNGK EYKCKVSNKA LAAPIEKTIS KAKGQPREPQ VYTLPPSREEMTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRWQQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 113) HCIgG1 (L234A, L235A) mutant constant region amino acid sequence (EU Numbering)ASTKGPSVFP LAPSSKSTSG GTAALGCLVK DYFPEPVTVS WNSGALTSGV HTFPAVLQSSGLYSLSSVVT VPSSSLGTQT YICNVNHKPS NTKVDKRVEP KSCDKTHTCP PCPAPEAAGGPSVFLFPPKP KDTLMISRTP EVTCVVVDVS HEDPEVKFNW YVDGVEVHNA KTKPREEQYNSTYRVVSVLT VLHQDWLNGK EYKCKVSNKA LPAPIEKTIS KAKGQPREPQ VYTLPPSREEMTKNQVSLTC LVKGFYPSDI AVEWESNGQP ENNYKTTPPV LDSDGSFFLY SKLTVDKSRWQQGNVFSCSV MHEALHNHYT QKSLSLSPGK (SEQ ID NO: 114)

TABLE 6 Selected therapeutic agents that can be administered incombination with the anti-TIM- 3 antibody molecules, e.g., as a singleagent or in combination with other immunomodulators described herein.Each publication listed in this Table is herein incorporated byreference in its entirety, including all structural formulae therein.Patents/Patent Compound Generic Name Application No. Tradename CompoundStructure Publications A1 Sotrastaurin

EP 1682103 US 2007/142401 WO 2005/039549 A2 Nilotinib HCl monohydrateTASIGNA ®

WO 2004/005281 U.S. Pat. No. 7,169,791 A3

WO 2010/060937 WO 2004/072051 EP 1611112 U.S. Pat. No. 8,450,310 A4Dactolisib

WO 2006/122806 A5

U.S. Pat. No. 8,552,002 A6 Buparlisib

WO 2007/084786 A7

WO 2009/141386 US 2010/0105667 A8

WO 2010/029082 A9 CYP17 inhibitor WO 2010/149755 U.S. Pat. No. 8,263,635B2 EP 2448903 B1 A10

WO 2011/076786 A11 Deferasirox EXJADE ®

WO 1997/049395 A12 Letrozole FEMARA ®

U.S. Pat. No. 4,978,672 A13

WO 2013/124826 US 2013/0225574 A14

WO 2013/111105 A15

WO 2005/073224 A16 Imatinib mesylate GLEEVEC ®

  Mesylate WO 1999/003854 A17

  Dihydrochloric salt EP 2099447 U.S. Pat. No. 7,767,675 U.S. Pat. No.8,420,645 A18 Ruxolitinib Phosphate JAKAFI ®

  H₃PO₄ WO 2007/070514 EP 2474545 U.S. Pat. No. 7,598,257 WO 2014/018632A19 Panobinostat

WO 2014/172493 WO 2002/022577 EP 1870399 A20 Osilodrostat

WO 2007/024945 A21

WO 2008/016893 EP 2051990 U.S. Pat. No. 8,546,336 A22 Sonidegibphosphate

WO 2007/131201 EP 2021328 U.S. Pat. No. 8,178,563 A23 ceritinibZYKADIA ™

WO 2008/073687 U.S. Pat. No. 8,039,479 A24

U.S. Pat. No. 8,415,355 U.S. Pat. No. 8,685,980 A25

WO 2010/007120 A26 Human monoclonal antibody to PRLR U.S. Pat. No.7,867,493 A27

WO 2010/026124 EP 2344474 US 2010/0056576 WO2008/106692 A28

WO 2010/101849 A29 Encorafenib

WO 2011/025927 A30

WO 2011/101409 A31 Human monoclonal antibody to HER3 WO 2012/022814 EP2606070 U.S. Pat. No. 8,735,551 A32 Antibody Drug Conjugate (ADC) WO2014/160160 Ab: 12425 (see Table 1, paragraph [00191]) Linker: SMCC (seeparagraph [00117] Payload: DM1 (see paragraph [00111] See also Claim 29A33 Monoclonal antibody or Fab to M-CSF WO 2004/045532 A34 Binimetinib

WO 2003/077914 A35 Midostaurin

WO 2003/037347 EP 1441737 US 2012/252785 A36 Everolimus AFINITOR ®

WO 2014/085318 A37

WO 2007/030377 U.S. Pat. No. 7,482,367 A38 Pasireotide diaspartateSIGNIFOR ®

WO2002/010192 U.S. Pat. No. 7,473,761 A39 Dovitinib

WO 2009/115562 U.S. Pat. No. 8,563,556 A40

WO 2013/184757 A41

WO 2006/122806 A42

WO 2008/173687 U.S. Pat. No. 8,372,858 A43

WO 2010/002655 U.S. Pat. No. 8,519,129 A44

WO 2010/002655 U.S. Pat. No. 8,519,129 A45

WO 2010/002655 A46 Valspodar AMDRAY ™

EP 296122 A47 Vatalanib succinate

  succinate WO 98/35958 A48 IDH inhibitor WO2014/141104 A49 BCR-ABLinhibitor WO2013/171639 WO2013/171640 WO2013/171641 WO2013/171642 A50cRAF inhibitor WO 2014/151616 A51 ERK1/2 ATP competitive inhibitorPCT/US2014/062913

-   Table 7. See Examples.-   Table 8. See Examples.-   Table 9. See Examples.-   Table 10. See Examples.-   Table 11. See Examples.-   Table 12. See Examples.-   Table 13. See Examples.-   Table 14. See Examples.-   Table 15. See Examples.-   Table 16. See Examples.

Examples Example 1 Characterization of ABTIM3 and Other Anti-TIM-3Antibodies

Panels of anti-TIM-3 antibodies were assayed for binding to TIM-3expressing cells. The dissociation constants (K_(D)) of two suchantibodies, ABTIM3 and anti-TIM-3 #2, as measured by surface plasmonresonance, is summarized in FIG. 2A. In FIGS. 2B and 2C, the binding ofvarious anti-TIM-3 antibodies, including ABTIM3, to cells transfectedwith human TIM-3 was measured using flow cytometry. Next, threeantibodies, ABTIM3, anti-TIM-3 #2, and anti-TIM-3 #3, and a controlantibody were assayed for the ability to bind cynomolgus TIM-3 in cellstransfected with cynoTIM-3. FIG. 2D shows that ABTIM3 has the strongestaffinity for cynomolgus TIM-3 out of the three antibodies tested. FIG.2E tests seven anti-human-TIM-3 antibodies for the ability to bindcynomolgus TIM-3, and shows that ABTIM3 binds with the highest affinity.Overall, the experiments indicate that ABTIM3 has strong (sub-nanomolar)affinity for both human and cynomolgus TIM-3.

The ability of three anti-TIM-3 antibodies, including ABTIM3, to bind tohuman TIM-4 expressed in CHO cells and murine TIM-3 expressed in cellswas also measured by flow cytometry. Human TIM-3 has about 23% sequenceidentity with human TIM-4. Murine TIM-3 has about 66% sequence identitywith human TIM-3 and 64% sequence identity with cynomolgus TIM-3. Theresults from these assays show that ABTIM3 does not bind to human TIM-4.ABTIM3 is also not cross-reactive with murine TIM-3. Taken together withthe binding assay results described above, ABTIM3 antibody is specificfor human and cynomolgus TIM-3.

In a cross-blocking experiment, ABTIM3 was shown to cross-blockanti-TIM-3 #2, suggesting that these antibodies bind to epitopes thatare near each other, and possibly overlap, although the two epitopes arenot necessarily identical.

The ability of TIM-3 antibodies, e.g., ABTIM3, to bind to activatedPBMCs expressing TIM-3 was also assessed. Whole human PBMCs from a donorwere stimulated for 10 days with platebound CD3/CD28 (1 μg/ml each), inthe presence of 10 ng/ml recombinant human IL-12. Cells were ficolled toremove dead cells and reactivated for three days with the same stimulus.Antibodies that bind to TIM-3, e.g., ABTIM3 and anti-TIM-3 #2, werecompared, and anti-PD-1, anti-PD-L1, and anti-LAG-3 antibodies, andmouse IgG1 were used as control antibodies. Cells were incubated withthe antibodies at various concentrations from 0.001 to 100 μg/ml, andthe antibody binding to the cells was analyzed by flow cytometry. Cellswere gated for CD4 or CD8 positive populations, and mean fluorescenceintensity (MFI) for each antibody and concentration was plotted on agraph. Dissociation constant (K_(D)) values were then calculated. Theresults from the assays are shown in Table 7 below.

TABLE 7 K_(D) values for anti-TIM-3 binding to activated PBMCs CD4 gatedPBMCs CD8 gated PBMCs Antibody K_(D) K_(D) ABTIM3 0.29 nM 0.30 nMAnti-TIM-3 #2 2.84 nM 3.14 nM Anti-PD-L1 control 0.20 nM 0.30 nMAnti-LAG-3 control 2.33 nM Anti-PD-1 control 22.8 nM 85.9 nM

These results demonstrate that ABTIM3 was able to bind to TIM-3expressed on activated PBMCs.

Example 2 Domain Analysis of Anti-TIM-3 Antibody Binding to TIM-3

TIM-3 has an extracellular IgV domain and a mucin domain. The regions ofTIM-3 bound by each of five antibodies was determined using arecombinant construct that replaced the IgV domain of TIM-3 with the IgVdomain of PD-1, and this construct is depicted in FIG. 3A. FIG. 3B showsthat the anti-TIM-3 monoclonal antibody (anti-TIM-3 #3), and twoanti-PD-L1 control monoclonal antibodies (anti-PD-L1 #1 and #2), bind tothe chimeric protein of FIG. 3A, while anti-TIM-3 #2 and ABTIM3 do notsubstantially bind. This result suggests that the anti-TIM-3 monoclonalantibodies anti-TIM-3 #2 and ABTIM3 bind to the IgV domain of TIM-3,while anti-TIM-3 #3 binds to the mucin domain of TIM-3. The dissociationconstant (K_(D)) values were calculated for each tested antibody for therecombinant construct are shown in Table 8.

TABLE 8 K_(D) values for binding to PD-L1 IgV/TIM-3 mucin constructAntibody Antigen K_(D) Anti-PD-L1 #1 PD-L1 IgV domain 0.52 nM Anti-PD-L1#2 PD-L1 IgV domain 0.38 nM Anti-TIM-3 #3 TIM-3 mucin domain 2.71 nMAnti-TIM-3 #2 TIM-3 No binding to the chimeric protein ABTIM3 TIM-3 Nobinding to the chimeric protein

Example 3 TIM-3 Binding to PtdSer is Blocked by Anti-TIM-3 Antibodies

TIM-3 binds to PtdSer (phosphatidylserine), a lipid that is typicallypresent on the surface of apoptotic cells and not normal cells.Anti-CD95-treated WR19L(Fas) cells were cultured under conditions thatpromote PtdSer accumulation on the cell surface (flipping of PtdSer fromthe inner membrane to external exposure upon induction of apoptosis).TIM-3-Ig (huTIM-3 extracellular domain fused to an Ig Fc region) wasadded to the cells, and binding of TIM-3-Ig to the cells was assayed inthe presence of various antibodies. As shown in FIG. 4, severalanti-TIM-3 mAbs, including ABTIM3, anti-TIM-3#5, and anti-TIM-3 #2,inhibit the binding of TIM-3 to PtdSer.

Example 4 IFN-Gamma Secretion of CD4+ Cells is Enhanced by Anti-TIM-3Antibodies

The ability of four antibodies to enhance IFN-gamma secretion andproliferation of IL-12 stimulated CD4+ cells was assayed. This assayused the observation that a high dose of IL-12 induces expression ofTIM-3 and yields an exhausted phenotype in T cells (see Yang et al., J.Clin. Invest. 122:4 p1271 2012). FIG. 5A shows four panels, each ofwhich indicates the results of an experiment where cells were exposed toan antibody selected from ABTIM3, anti-TIM-3 #2, mIgG1, and anti-PD-L1antibody (anti-PD-L1 control). After PMA/ionomycin restimulation andfixation and permeabilization of cells, the resulting IFN-gamma levelswere measured by flow cytometry (y axis) and proliferation was measuredby CFSE fluorescence (x axis). FIG. 5B quantifies IFN-gamma expressionin cells exposed to these four antibodies. From left to right, the barsin FIG. 5B correspond to antibodies ABTIM3, anti-TIM-3 #2, anti-PD-L1control, and mIgG1.

Example 5 TIM-3 Blockade Enhances In Vitro Functional Activity

5.1 TIM-3 Blockade Enhances In Vitro Cytotoxic Activity of Purified NKCells

TIM-3 is highly expressed endogenously on NK (natural killer) cells; itsexpression is further induced on activated NK cells. TIM-3 may act torestrain NK cell function, as do other inhibitory receptors. See Ndhlovuet al., Blood 119:3734, 2012, and Silva et al., Cancer Immunol Res2:410, 2014. Accordingly, the ability of ABTIM3 and other anti-TIM-3antibodies to enhance NK cell cytotoxic activity was assayed.

In this assay, NK cells were purified from whole blood by negative beadselection and then incubated with antibody (10 μg/mL) at 37° C. After 1hour, target K562 cells were added. After a 4-hour incubation at 37° C.,the percent of K562 cell killing was measured. Antibody ABTIM3 resultedin elevated levels of K523 cell killing relative to anti-TIM-3 #2 or theisotype control.

5.2 TIM-3 Blockade Increases Proliferation from Autologous T-DCCo-Cultures

TIM-3 can be expressed on dendritic cells (DCs) and T cells. Naïve Tcells and dendritic cells were isolated from donor samples. Naïve Tcells and conventional DCs were cocultured for four days in the presenceof anti-CD3/CD28. ABTIM3 was added at varying doses, 0.01 μg/mL, 0.1μg/mL, 1 μg/mL, 5 μg/mL, and 25 μg/mL, to the co-culture. Cellproliferation was detected by a CFSE proliferation assay, which relieson dilution of CFSE staining to detect proliferating cells.

As shown in FIG. 22, the presence of ABTIM3 at every tested dosageresulted in an increase in proliferating cells, as represented byCFSE-diluted cells, compared to no antibody and the mouse isotype (IgG1)control.

Example 6 Characterization of Humanized Anti-TIM-3 Antibody

6.1 Generation of Humanized Anti-TIM-3 Antibodies

The murine anti-TIM-3 antibody ABTIM3 was humanized by grafting theCDRs, e.g., provided in Table 3, to human IgG4 constant region, with astabilized hinge region containing the S228P mutation. Additionalmodifications were made to the CDR2 of the heavy chain by mutating theputative deamidation site from N at position 6 of HCDR (Kabat), orposition 4 of the HCDR2 (Chothia) to S or Q to remove the deamidationsite. Other modifications included using alternative frameworks. Theunique heavy chains and light chains combined in various combinations togenerate a small library of unique humanized mAbs.

6.2 Binding Assays

The binding capability of the humanized mAbs generated were tested bycompetition binding with the parent murine anti-TIM-3 antibody in afluorescence-activated sorting assay. A representative graph depictingthe results from the FACs-based competition assay comparing the bindingbetween the parent murine anti-TIM-3 antibody and 4 humanized anti-TIM-3antibodies (ABTIM3-hum01, ABTIM3-hum04, ABTIM3-hum07, and ABTIM3-hum08),and hIgG4 control is shown in FIG. 7.

The results from multiple surface plasmon resonance Biacore bindingassays for a panel of humanized anti-TIM-3 antibodies are summarized inTable 9.

TABLE 9 Biacore K_(D) values for a panel of anti-TIM-3 antibodies KD(nM) KD (nM) KD (nM) KD (nM) Clone 4.7.14 4.29.14 5.1.14 5.30.14ABTIM3-hum02 0.308 0.269 0.174 ABTIM3-hum03 0.351 0.16 0.314ABTIM3-hum05 0.313 0.279 0.332 ABTIM3-hum06 0.498 0.214 0.364ABTIM3-hum09 0.161 ABTIM3-hum10 0.107 ABTIM3-hum11 0.194 ABTIM3-hum120.355 ABTIM-hum01 0.23 ABTIM-hum04 0.172 ABTIM3-hum01 0.103 0.114 0.193ABTIM3-hum07 0.135 0.199 0.196 ABTIM3-hum08 0.123 0.309 0.175ABTIM3-hum04 0.216All of the tested humanized mAbs were demonstrated to have relativelythe same affinity as each other and the parent murine anti-TIM-3antibody, within 0.1-0.5 nM K_(D).

6.3 Binding to TIM-3 Expressing Cells

The humanized anti-TIM-3 antibodies were assayed for binding to TIM-3expressing cells using fluorescence activated cell sorting and Biacoreassays, described in Example 1. In FIG. 8A, the binding of varioushumanized anti-TIM-3 antibodies to cells transfected with human TIM-3was measured using flow cytometry. ABTIM3 was used as a positivecontrol. Negative controls include hIgG4, goat anti-human, and goatanti-mouse secondary Ab-FITC. The results from the flow cytometrycompetition assay were used to determine the dissociation constant(K_(D)) for cells expressing human TIM-3, as shown in Table 10 below.

TABLE 10 K_(D) values for binding to cells expressing huTIM-3. AntibodyK_(D) (nM) ABTIM3-hum03 0.887 ABTIM3-hum11 0.906 ABTIM3-hum21 0.917ABTIM3 1.04

A competition binding assay was also performed to assess binding of thehumanized anti-TIM-3 antibodies, ABTIM3-hum03 and ABTIM3-hum11, to cellsexpressing human TIM-3, while in the presence of the parental murineantibody, ABTIM3. As shown in FIG. 8B, the humanized anti-TIM-3antibodies competed with ABTIM3.

The K_(D) values for two humanized anti-TIM-3 antibodies for recombinantTIM-3-Ig fusion proteins were assayed by surface plasmon resonance in aBiacore assay, as shown in Table 11.

TABLE 11 Biacore K_(D) values for TIM-3-Ig cynoTIM- huTIM- mTIM- ratTIM-3/Fc 3/his 3/his 3/Fc ABTIM3-hum03 KD(M) 1.04E−09 1.24E−10 KD(M)3.89E−09 1.84E−10 5.10E−08 KD(M) 3.08E−09 7.58E−11 Mean 2.67E−091.28E−10 KD(M) ABTIM3-hum11 KD(M) 1.24E−09 1.55E−10 KD(M) 3.14E−092.26E−10 KD(M) 5.04E−09 1.09E−10 2.97E−07 Mean 3.14E−09 1.63E−10 KD(M)

These results show that the humanized TIM-3 antibodies have similarbinding affinity with human and cynomolgus proteins. The humanized TIM-3antibodies showed very weak binding affinity to rat TIM-3/Fc protein, inthe order of 1/1000 compared to the binding affinity with huTIM-3/Fc.

Example 7 X-Ray Crystal Structure of the Human TIM-3/ABTIM3-hum21 FabComplex

The crystal structure of a human TIM-3 (IgV domain, SEQ ID NO: 220,Table 12) bound to the Fab fragment of a humanized anti-TIM-3 antibodyABTIM3-hum21 (SEQ ID NO: 221 and 222, Table 12) was determined. Asdetailed below, TIM-3 was co-expressed with MGB220 Fab in mammaliancells to produce purified complex. Protein crystallography was thenemployed to generate atomic resolution data for TIM-3 bound toABTIM3-hum21 Fab to define the epitope. ABTIM3-hum21, a humanizedantibody from a parental murine antibody, comprises an IgG1 frameworkand the variable heavy chain of SEQ ID NO: 84, and the variable lightchain of SEQ ID NO: 88. ABTIM3-hum21 differs by only one amino acid inheavy chain CDR2 from other humanized anti-TIM antibodies describedherein and this different amino acid (Gln55) is far away (>6 Å) from theepitope and thus would not change antigen binding, which indicates thatthe crystal structure results obtained are applicable to the otherhumanized antibodies described herein.

7.1 Protein Production

The sequences of TIM-3 and ABTIM3-hum21 Fab produced for crystallographyare shown in Table 12. The construct of TIM-3 comprises residues 22 to130 (underlined) of human TIM-3 (UniProt identifier Q8TDQ0, SEQ ID NO:129), along with N- and C-terminal residues from recombinant expressionvector (shown in lower case letters, SEQ ID NO: 130). The N-terminalsignal sequence from mouse IgG kappa light chain was used for secretedexpression of TIM-3 and was cleaved during expression, leaving intactN-terminus of TIM-3. The C-terminus of TIM-3 was fused with a 6×His tag(SEQ ID NO: 133) for purification. For ABTIM3-hum21 Fab, the sequencesof heavy (SEQ ID NO: 131) and light (SEQ ID NO: 132) chains are shown.

TABLE 12 Amino acid sequences used for crystal structure determinationSEQ ID Construct Amino acid sequence NO Human TIM-3MFSHLPFDCVLLLLLLLLTRSSEVEYRAEVGQNAYLPCF 129 (Q8TDQ0)YTPAAPGNLVPVCWGKGACPVFECGNVVLRTDERDVNYWTSRYWLNGDFRKGDVSLTIENVTLADSGIYCCRIQIPGIMNDEKFNLKLVIKPAKVTPAPTRQRDFTAAFPRMLTTRGHGPAETQTLGSLPDINLTQISTLANELRDSRLANDLRDSGATIRIGIYIGAGICAGLALALIFGALIFKWYSHSKEKIQNLSLISLANLPPSGLANAVAEGIRSEENIYTIEENVYEVEEPNE YYCYVSSRQQPSQPLGCRFAMPHuman TIM-3 metdtlllwvlllwvpgstgSEVEYRAEVGQNAYLPCFYTPAAPGN 130expression LVPVCWGKGACPVFECGNVVLRTDERDVNYWTSRYWL constructNGDFRKGDVSLTIENVTLADSGIYCCRIQIPGIMNDEKFN LKLVIKhhhhhh ABTIM3-QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWV 131 hum21 FabRQAPGQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKS heavy chainTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH ABTIM3-DIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQW 132 hum21 Fab lightYQQKPGQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTI chainSSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC

TIM-3 was co-expressed with ABTIM3-hum21 Fab in Expi293® cells toproduce complex for crystallography. In detail, 0.3 mg of plasmidencoding TIM-3 was mixed with 0.15 mg of plasmid encoding the heavychain of ABTIM3-hum21 Fab and 0.15 mg of plasmid encoding the lightchain of ABTIM3-hum21 Fab, diluted into 30 mls of Opti-MEM® I medium(Life Technologies), and incubated with 1.5 mgs of Polyethylenimine(Polysciences) in 30 mls of the same medium for 30 min. The mixture wasthen added into 0.6 L of Expi293® cells growing in suspension inExpi293® Expression medium (Life Technologies) at 2 million cells/ml at37° C. with 8% of CO₂ for transfection. After 5 days, the mediumcontaining TIM-3/ABTIM3-HUM21 Fab complex was harvested bycentrifugation. Five mls of Ni-NTA resin was added into the medium andkept stirring at 4° C. overnight. The next day the beads were packedinto a gravity column and washed with 25 mM Hepes pH 7.4, 150 mM NaCl(HBS) supplemented with 20 mM of imidazole. The complex was eluted with3 column volumes (CV) of HBS with 500 mM of imidazole, and then dialyzedin HBS at 4° C. overnight. The next day, the complex was incubated with1/10 (w/w) of PNGaseF (purified in-house) at 37° C. overnight to removeN-linked glycosylation. After deglycosylation, the mixture was boundback to 5 mls of Ni-NTA resin, washed with HBS to remove PNGaseF andeluted with HBS plus 500 mM of imidazole. The eluate was thenconcentrated and loaded onto HiLoad 16/600 Superdex 75 PG (GEHealthcare) size exclusion column equilibrated in HBS. Peak fractionscontaining purified TIM-3/ABTIM3-hum21 Fab complex were analyzed bySDS-PAGE, pooled and concentrated for crystallization.

7.2 Crystallization and Structure Determination

TIM-3/ABTIM3-hum21 Fab complex was concentrated to 12.5 mg/ml,centrifuged at 20,000 g for 10 min, and screened for crystallization.Crystals for data collection were grown by hanging drop vapor diffusionat 20° C. In detail, 0.1 μl of the TIM-3/ABTIM3-hum21 Fab complex wasmixed with 0.1 μl of reservoir solution containing 0.04 M potassiumphosphate monobasic, 16% (w/v) PEG 8000 and 20% (v/v) Glycerol. The dropwas then equilibrated against 45 μl of the same reservoir solution.Before data collection, the crystals were flash cooled in liquidnitrogen.

Diffraction data were collected at beamline 17-ID at the Advanced PhotonSource (Argonne National Laboratory, USA), and processed using Autoproc(version 1.1.5, Global Phasing, LTD). The data of TIM-3/ABTIM3-hum21 Fabwas processed to 2.0 Å in space group P2₁ with cell dimensions a=84.3 Å,b=93.0 Å, c=85.3 Å, alpha=90°, beta=114°, and gamma=90°. The structureof the complex was solved by molecular replacement using Phaser (version2.5.5, McCoy et al., (2007) J. Appl. Cryst. 40:658-674) with structuresof mouse TIM-3 (PDB ID: 3KAA) and a Fab (in-house structure) as searchmodels. The final model was built in COOT (version 0.6 pre, Emsley &Cowtan (2004) Acta Cryst. D 60:2126-2132) and refined using Phenix(version 1.9, Afonine et al., (2012) Acta Cryst. D 68:352-367). TheR_(work) and R_(free) values were 17.5% and 22.1%, respectively; and theroot-mean-square (r.m.s) deviation values of bond lengths and bondangles are 0.007 Å and 1.1°, respectively.

Epitope was defined as residues of TIM-3 that contain atoms within 5 Åto any atom in ABTIM3-hum21 Fab, identified by CONTACT in CCP4 programsuite (version 6.2.0, Winn et al., (2011) Acta. Cryst. D 67:235-242) andlisted in Table 13. There are 2 copies of TIM-3/ABTIM3-hum21 Fabcomplexes in the asymmetric unit (the smallest unique unit in thecrystal), only those antibody-contacting residues that are common inboth copies are listed as epitope residues.

7.3 Epitope of ABTIM3-hum21 on TIM-3

The crystal structure of the TIM-3/ABTIM3-hum21 Fab complex was used toidentify the epitope of ABTIM3-hum21 on TIM-3. The interaction surfaceon TIM-3 by ABTIM3-hum21 was formed by several continuous anddiscontinuous (i.e. noncontiguous) sequences: namely residues Val24,Glu25, Tyr26, Phe39, Tyr40, Thr41, Gly56, Ala57, Cys58, Pro59, Val60,Phe61, Ser105, Gly106, Ile107, Asn119, Asp120, Glu121, Lys122, Phe123,Asn124, Leu125, Lys126, Leu127, and Val128 as detailed in Table 13.These residues form the exemplary three-dimensional conformationalepitope that is recognized by ABTIM3-hum21 (FIG. 9).

TABLE 13 Interactions between human TIM-3 and ABTIM3-hum21. TIM-3residues are numbered as in UniProt entry Q8TDQ0 (SEQ ID NO: 219). Theantibody residues are numbered based upon their linear amino acidsequence (SEQ ID NO: 221 and 222) and corresponding chains are labeled(“H” for heavy chain, “L” for light chain). TIM-3 residues shown herehave at least one atom within 5 Å to an atom in ABTIM3-hum21, to accountfor potential water mediated interactions. TIM-3 ABTIM3-hum21 Amino acidNumber Amino acid Number Chain Val 24 Ala 102 H Asp 98 L Glu 25 Tyr 31 LArg 96 L Tyr 26 Tyr 31 L Phe 39 Ser 31 H Tyr 52 H Tyr 40 Ser 31 H Thr 28H Thr 41 Thr 28 H Gly 56 Thr 34 L Ala 57 Phe 103 H Thr 34 L Asn 57 L Tyr53 L Ala 54 L Cys 58 Tyr 53 L Asn 57 L Pro 59 Asn 57 L Tyr 53 L Val 60Asn 57 L Tyr 53 L Val 58 L Ser 60 L Glu 59 L Phe 61 Ser 60 L Ser 105 Tyr32 L Gly 106 Tyr 31 L Tyr 32 L Ile 107 Phe 103 H Thr 34 L Tyr 31 L Leu36 L Asn 119 Ser 60 L Asp 120 Tyr 32 H Glu 121 Tyr 32 H Thr 28 H Lys 122Tyr 32 H Gly 100 H Tyr 53 L Glu 59 L Phe 123 Gly 100 H Gly 101 H Tyr 32H Asn 124 Phe 103 H Ala 102 H Pro 104 H Tyr 53 L Leu 125 Ala 102 H Lys126 Ala 102 H Tyr 31 L Leu 36 L Ser 95 L Lys 97 L Leu 127 Tyr 31 L Val128 Tyr 31 L Tyr 32 L

7.4 ABTIM3-hum21 v.s. TIM-3 Ligands

The identification of the epitope of TIM-3 recognized by the anti-TIM-3antibody indicates that binding of some of the TIM-3 ligands may bedisrupted by antibody binding. The known ligands of TIM-3 includeCEACAM-1, phosphatidylserine (PtdSer), HMGB1, and Galectin-9 (Gal-9).

With respect to CEACAM-1, a recent study has showed that CEACAM-1 is aligand for TIM-3 required for its ability to mediate T-cell inhibition,and this interaction has a crucial role in regulating autoimmunity andanti-tumour immunity (Huang et al., (2014) Nature). The same study alsoidentified, both biochemically and structurally, the crucial amino acidresidues of TIM-3 mediating its binding to CEACAM-1 (FIG. 10A). TheABTIM3-hum21 epitope on TIM-3 overlaps with these CEACAM-1-bindingresidues (FIG. 10A), including C58, N119 and K122. For example, K122forms hydrogen bond N42 of CEACAM-1, but is complete blocked byABTIM3-hum21 (FIG. 10B). Superimposition of the crystal structuresobtained from the TIM-3/ABTIM3-hum21 Fab and the TIM-3/CEACAM-1 (PDB ID:4QYC) complexes results in a significant clash between ABTIM3-hum21 andCEACAM-1 (FIG. 10C). Altogether, these data suggests that ABTIM3-hum21disrupts CEACAM-1 binding.

With respect to PtdSer, the FG loop and CC′ loop of TIM-3 form a pocket(often referred to as the metal ion-dependent ligand binding site(MILIBS)) that has been shown by crystal structure to bind Ca²⁺ andPtdSer simultaneously (DeKruyff, et al., (2010) J Immunol.184(4):1918-1930). This binding is thought to help TIM-3-expressingcells engage and penetrate the membrane of apoptotic cells (whichdisplays PtdSer) for engulfment. The crystal structure ofTIM-3/ABTIM3-hum21 Fab indicates that ABTIM3-hum21 binds thePtdSer-binding loops of the human TIM-3 IgV domain; and the attackingangle of the antibody suggests it will prevent PtdSer-mediated membranepenetration of TIM-3 (FIG. 11).

With respect to HMGB1, it has been reported to interact with TIM-3 tohelp tumor-associated dendritic cells suppress nucleic acid-mediatedinnate immune response (Chiba et al., (2012) Nat. Immunol.13(9):832-842). The amino acid residue at position 62 of TIM-3 (Q inmouse, E in human TIM-3) has been shown to be important for mouse HMGB1binding to mouse TIM-3. E62 is not present in the ABTIM3-hum21 epitope,though it is very close to the two epitope residues V60 and F61, thusthere is a chance that ABTIM3-hum21 can block HMGB1 binding depending onthe attacking angle of HMGB1 to TIM-3.

With respect to Gal-9, it has been shown to bind mouse TIM-3 tonegatively regulate Th1-immune response (Zhu et al., (2005) Nat.Immunol. 6(12):1245-1252). However, it has also been reported that humanTIM-3 on T cells does not act as a receptor for Gal-9 (Leitner et al.,(2013) PLoS Pathog. 9(3):e1003253). From the crystal structure of humanTIM-3/ABTIM3-hum21 Fab, half of the proposed Gal-9 binding site in mouseTIM-3 is not conserved in human TIM-3 (N74 and N90 in mouse TIM-3 becomeD74 and R89 in human TIM-3), i.e. this half-site in human TIM-3 will notbe able to bind Gal-9. The left-over half site (N33 and N99 in humanTIM-3) is conserved but is far away from the ABTIM3-hum21 epitope onTIM-3 (FIG. 9A). Therefore, even if Gal-9 is a ligand of human TIM-3,ABTIM3-hum21 will not disrupt the binding of Gal-9 to human TIM-3.

7.5 Hydrogen-Deuterium Exchange Experimental Setup

HDx/MS experiments were performed using methods similar to thosedescribed in the literature (Chalmers et al., (2006) Anal. Chem.78(4):1005-1014). The experiments were performed on a Waters HDx/MSplatform, which includes a LEAP autosampler, nanoACQUITY UPLC and SynaptG2 mass spectrometer. The deuterium buffer used to label the proteinbackbone of human TIM-3 (aa22-135; SEQ ID NO: 139) was 25 mM HEPES, 150mM NaCl, 5 mM CaCl₂ pH7.4 with deterium; the overall percentage ofdeuterium in the solution was 94.2%. For human TIM-3 (aa22-135)deuterium labeling experiments in the absence of antibody, 300 pmol ofhuman TIM-3 (aa22-135), volume of 7.7 μl, was diluted using 100 μl ofthe deuterium buffer in a chilled tube and incubated for 15 minutes on arotator at 4° C. The labeling reaction was then quenched with 100 μl ofchilled quench buffer at 2° C. for five minutes followed by injectedonto the LC-MS system for automated pepsin digestion and peptideanalysis.

For human TIM-3 (aa22-135) deuterium labeling experiments in thepresence of antibodies, 400 pmol of ABTIM3-hum03 or ABTIM3-hum11 wasfirst immobilized on Thermo Protein G Plus beads and cross-linked usingdisuccinimidyl suberate (DSS). To perform the labeling experiments, theantibody beads (containing 400 pmol antibody) were incubated with 300pmol human TIM-3 (aa22-135) for 25 minutes at 4° C. After 25 minutes thebeads were washed with 200 μl of HEPES buffer. Then 200 μl of chilleddeuterium buffer (87.5% deuterium) was added and the complex wasincubated for 15 minutes at 4° C. After 15 minutes, the deuterium bufferwas spun out and the labeling reaction was quenched with 200 μl ofchilled quench buffer on ice for 4 minutes. After spinning the samplefor 30 seconds in a centrifuge, the quenched solution was injected ontothe LC-MS system for automated pepsin digestion and peptide analysis.

All deuterium exchange experiments were quenched using 1 M TCEP and 6 Murea (pH 2.6). After quenching, the exchanged antigen was subjected toon-line pepsin digestion using a Poroszyme Immobilized Pepsin column(2.1×30 mm) at 12° C. followed by trapping on a Waters Vanguard HSS T3trapping column. Peptides were eluted from the trapping column andseparated on a Waters BEH C18 1×100 mm column (maintained at 1° C.) at aflow rate of 40 μl/min using a binary 8.4 minute gradient of 2 to 35% B(mobile phase A was 99.9% water and 0.1% formic acid; mobile phase B was99.9% acetonitrile and 0.1% formic acid).

7.6 Hydrogen-Deuterium Exchange Results

For human TIM-3 93% of the sequence was monitored by deuterium exchangeas shown in FIG. 18. In this figure each bar represents a peptide thatis monitored in all deuterium exchange experiments. For differentialexperiments between antibody bound and unbound states it is informativeto examine the difference in deuterium uptake between the two states. InFIG. 19 a negative value indicates that the TIM-3-antibody complexundergoes less deuterium uptake relative to TIM-3 alone. A decrease indeuterium uptake can be due to protection of the region fromexchangeable deuterium or stabilization of the hydrogen bonding network.In contrast, a positive value indicates that the complex undergoes moredeuterium uptake relative to TIM-3 alone. An increase in deuteriumuptake can be due to destabilization of hydrogen bonding networks (i.e.localized unfolding of the protein).

ABTIM3-hum03 shares identical CDRs with ABTIM3-hum11 except thatABTIM3-hum03 has a glutamine at position 55 in HCDR2 while ABTIM3-hum11has an asparagine at position 55 in HCDR2. ABTIM3-hum03 shares the sameCDR regions with ABTIM3-hum21. One expects these antiboides to have thesame epitope on TIM-3. From FIG. 19 one observes that ABTIM3-hum03 andABTIM3-hum11 exhihit the same protection profile which is consistentwith the two antibodies sharing the same epitope. Closer examination ofFIG. 19 reveals that when TIM-3 is complexed with either of the twoantibodies that many regions of TIM-3 undero significant protection,typically defined as proection less than or equal to −0.5 Da (Houde etal. (2010) J. Pharma. Sci. 100(6): 2071-2086). The observation of broadprotection suggests that binding of either of the two antibodies to theTIM-3 antigen cause a broad based stabilization of hydrogen bondingnetworks in TIM-3. This broad protection is in addition to theprotection that results from solvent sheilding of the epitope at theantibody-antigen interface. Given the significant amount of broadprotection, it is useful to rank order the most protected regions ofTIM-3 upon antibody binding to delineat the regions likely to beinvolved in the epitope. TIM-3 regions that are the most protected uponABTIM3-hum03 or ABTIM3-hum11 binding include the regions 23-25 (EVE),41-61 (TPAAPGNLVPVCWGKGACPVF, SEQ ID NO: 140), 73-77 (RDVNY, SEQ ID NO:141), and 121-127 (EKFNLKL, SEQ ID NO: 142). Comparing these protectedregions to the X-ray crstyal structure data summarized in Table 13 showsconsistent agreement indicating that the epitope determined by X-raycrystal structure is present in solution.

Example 8 TIM-3 Expression in Cancer

TIM-3 is expressed in various cancers. In this example, severaldifferent analysis methods were used to identify cancers with TIM-3expression in which therapeutic benefit could be achieved by ananti-TIM-3 antibody.

8.1 Immunohistochemical Staining of Tumors

ABTIM-3 was used to stain various tumor tissues. TIM-3 tumor expressionwas identified in esophageal squamous cell carcinoma, primary andmetastatic renal cell carcinoma, colorectal cancer, and leukemic stemcells in AML.

8.2 Expression Analysis in TCGA and ICGC Databases

Overall TIM-3 expression was compared in the The Cancer Genome Atlas(TCGA) database and the International Cancer Genome Consortium (ICGC)database. The following cancers were identified as among the highestexpressors of TIM-3: diffuse large B cell lymphoma (DLBCL), kidney renalclear cell carcinoma (KIRC), glioblastoma multiforme (GBM),nasopharyngeal carcinoma (NPC), lung adenocarcinoma (LUAD), kidney renalpapillary cell carcinoma (KIRP), mesothelioma (MESO), acute myeloidleukemia (AML), and in breast cancer, triple negative (TN)immunomodulatory (IM) subtype (FIG. 12).

Next, cancers were identified that were characterized by high TIM-3expression in conjunction with high expression of other immune cellmarkers. The other immune cell markers include: T cell marker CD3e, Tregulatory cell marker FoxP3, natural killer cell marker NKp30,macrophage marker CD68, and dendritic cell marker CD11c. As shown inFIG. 12, cancer indications with high expression of TIM-3 and the otherimmune cell marker were identified. quartile “High” expression wasquantified by 3^(rd) (or top 25%) expressors across more than 34,000cases. For TIM-3 and CD3e, the top indications were diffuse large B celllymphoma (DLBCL), nasopharyngeal carcinoma (NPC), and kidney renal clearcell carcinoma (KIRC). For TIM-3 and FoxP3, the top indications werediffuse large B cell lymphoma (DLBCL), nasopharyngeal carcinoma (NPC),and lung adenocarcinoma (LUAD). For TIM-3 and NKp30, the top indicationswere diffuse large B cell lymphoma (DLBCL), nasopharyngeal carcinoma(NPC), and acute myeloid leukemia (AML). For TIM-3 and CD68, the topindications were diffuse large B cell lymphoma (DLBCL), kidney renalclear cell carcinoma (KIRC), and kidney renal papillary cell carcinoma(KIRP). For TIM-3 and CD11c, the top indications were diffuse large Bcell lymphoma (DLBCL), mesothelioma (MESO) (though only a small samplewas assessed), and kidney renal papillary cell carcinoma (KIRP).

A comparison was also performed of the correlation between TIM-3 or PD-1to T cell associated or macrophage associated markers in the TCGAdatabase. The analysis revealed correlation between TIM-3 expression andboth T cell associated markers (e.g., ZAP70, CD3D, CD3G, CD8B, GZMH,GZMK, and ITK) and macrophage associated markers (e.g., LILRB4, MRC1,MSR1, SIGLEC1, TREM2, CD163, ITGAX, and ITGAM), however, TIM-3expression is more associated with macrophage markers, especiallyinhibitory receptors on macrophages (e.g., LILRB4). Expression of amacrophage signature, e.g., macrophage associated markers (e.g., LILRB4,MRC1, MSR1, SIGLEC1, TREM2, CD163, ITGAX, and ITGAM) was determined forvarious cancers and were organized for the highest expressors of themacrophage signature in FIG. 13. The cancer indications with highexpression of the macrophage signature are also the same indicationswith high expression of TIM-3.

Example 9 Patient Selection Based on PDL1/CD8/IFN-γ Status

For each of several types of cancer, samples from multiple patients weretested for PDL1/CD8/IFN-γ status. Each sample was classified as:triple-negative for PDL1/CD8/IFN-γ, single or double positive for thesemarkers, or triple-positive for these markers. FIG. 14 shows that inthis experiment, within a population of patients, the following types ofcancer are frequently triple-positive for PDL1/CD8/IFN-γ: Lung cancer(squamous), lung cancer (adenocarcinoma), head and neck cancer, cervicalcancer (squamous), stomach cancer, thyroid cancer, melanoma, andnasopharyngeal cancer. Patients having these types of cancer are goodcandidates for therapy with anti PD-1 antibodies and combinationtherapies as described herein. The likelihood of successful treatmentcan be further boosted by determining which patients are triple-positivefor PDL1/CD8/IFN-γ, and treating the triple-positive patients withanti-TIM-3 antibodies, alone or in combination with one or moreimmodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor), and/orcombination therapies, as described herein.

FIG. 15 shows that within a population of patients, the following typesof cancer are rarely triple positive for PDL1/CD8/IFN-γ: ER+ breastcancer and pancreatic cancer. Notably, even in cancers that aregenerally not positive for PDL1/CD8/IFN-γ, one can increase thelikelihood of successful treatment by determining which patients aretriple-positive for PDL1/CD8/IFN-γ, and treating the triple-positivepatients with anti-TIM-3 antibodies, alone or in combination with one ormore immodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor), and/orcombination therapies, as described herein.

FIG. 16 shows the proportion of breast cancer patients that are triplepositive for PDL1/CD8/IFN-γ. Considering breast cancer in general, theproportion of triple-positives is somewhat low. However, when onefocuses only on IM-TN breast cancer, it can be seen that a much largerpercentage of patients is triple positive for PDL1/CD8/IFN-γ. IM-TNbreast cancer is particularly difficult to treat with conventionaltherapies. The discovery that IM-TN breast cancer is oftentriple-positive for PDL1/CD8/IFN-γ opens up new avenues of therapy forthis cancer with anti-TIM-3 antibodies, alone or in combination with oneor more immodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor),and/or combination therapies, as described herein.

FIG. 17 shows the proportion of colon cancer patients that are triplepositive for PDL1/CD8/IFN-γ. Considering colon cancer in general, theproportion of triple-positive is somewhat low. However, when one focusesonly on MSI-high (high microsatellite instability) breast cancer, it canbe seen that a much larger percentage of patients is triple positive forPDL1/CD8/IFN-γ. MSI levels can be assayed using, e.g., commerciallyavailable PCR-based methods.

Gastric cancer samples were tested for levels of PDL1/CD8/IFN-γ (datanot shown). It was found that in MSI-high or EBV+ gastric cancers, about49% were positive for PDL1, and a high proportion of the PDL1-positivecells were triple positive for PDL1/CD8/IFN-γ. It was also found that aproportion of PDL1-positive cells and PDL1/CD8/IFN-γ positive cells werealso positive for PIK3CA. This finding suggests that these cancers maybe treated with an anti-TIM-3 antibody, alone or in combination with oneor more immodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor),optionally in combination with a PIK3 therapeutic.

MSI-high CRC samples were tested for a combination of markers (data notshown). It was found that in MSI-high CRC samples, a high proportion ofthe PDL1/CD8/IFN-γ samples are also positive for LAG-3, PD-1 (alsocalled PDCD1), RNF43, and BRAF. This finding suggests that these cancersmay be treated with an anti-TIM-3 antibody, optionally in combinationwith a therapeutic that targets one or more of LAG-3, PDCD1, RNF43, andBRAF.

Squamous cell lung cancers were tested for a combination of markers(data not shown). It was found that in squamous cell lung cancersamples, a high proportion of the PDL1/CD8/IFN-γ samples are alsopositive for LAG-3. This finding suggests that these cancers may betreated with an anti-TIM-3 antibody, optionally in combination with atherapeutic that targets LAG-3, e.g., a LAG-3 antibody.

Papillary thyroid cancers were tested for a combination of markersincluding the BRAF V600E mutation (data not shown). It was found that ahigh proportion of thyroid cancer samples that are positive for PDL1 arealso positive for BRAF V600E. This finding suggests that these cancersmay be treated with an anti-TIM-3 antibody, alone or in combination withone or more immodulators (e.g., a PD-1 inhibitor or a PD-L1 inhibitor),optionally in combination with a therapeutic that targets BRAF.

Example 10 Patient Selection Based on PD-L1 Status

To enable broad examination of cancer indications for PD-1/PD-L1 basedtherapies, we evaluated PD-L1 expression at both the protein and mRNAlevel in human cancers including both lung and hepatic tumors.

PD-L1 protein expression was evaluated in a set of formalin-fixedparaffin-embedded non-small cell lung (NSCLC) adenocarcinoma (ACA),NSCLC squamous cell carcinoma (SCC), and hepatocellular carcinoma (HCC)tumors by immunohistochemistry (IHC). PD-L1 expression was scoredsemi-quantitatively by a manual histo-score (H-score) methodology basedon staining intensity and percentage of positive tumor cells. In our IHCanalysis, PD-L1 positivity (PD-L1+) was defined as an H-score 20. Inparallel, PD-L1 mRNA expression data was examined from The Cancer GenomeAtlas (TCGA) in these same indications (503 NSCLC ACA, 489 NSCLC SCC,and 191 HCC) and analyzed by comparing the expression in matched normaltissues from TCGA.

With RNAseq analysis, data was calculated as log 2 (RPKM+0.1) after RSEMnormalization, utilizing OmicSoft RNASeq pipelines across TCGA tumorindications. The expression of PD-L1 is elevated in NSCLC ACA and SCC,relative to that in HCC. By overlaying the distributions and comparingthe expression levels across all indications in TCGA, we rankedoverexpression profiles for PD-L1 and found the TCGA HCC cohort to havemuch reduced PD-L1 mRNA levels, with a median level of −0.8 compared to1.3 for ACA and 1.5 for SCC, which amounts to more than a 2-fold changeof median level expression. With RNAseq, our analysis defines 50% ofNSCLC adenocarcinoma, 54% of NSCLC squamous cell carcinoma, and 6% ofHCC as high expressers for PD-L1.

Tumor cell PD-L1 protein expression was measured in 45 lungadenocarcinoma (ACA) samples, 47 lung squamous cell carcinoma (SCC)samples, and 36 hepatocellular carcinoma (HCC) samples. 16/45 (35.6%)lung ACA, 21/47 (44.7%) lung SCC were PD-L1 positive. In contrast, PD-L1positivity was seen in only 2/36 (5.6%) HCC samples.

In summary, with IHC and RNAseq analysis in large and independent humanNSCLC and HCC sample sets, we have found PD-L1 expression to be moreenriched in NSCLC than in HCC. Within NSCLC, there are comparablefindings between adenocarcinoma and squamous cell carcinomas.Importantly, amongst the large number of samples (128 for IHC and 1183for RNAseq) in the 3 indications, very good concordance is observedbetween protein- and mRNA-based analyses. Our finding thus establishesthe basis for large scale mRNA-based data mining in TCGA for indicationsand patient segments that may be enriched for responses to PD-1/PD-L1-and/or TIM-3 based immune therapies.

Example 11 Competition Assays Indicate Humanized Anti-TIM3 AntibodiesBind to a Similar Epitope

As described above, the epitope of TIM-3 recognized by ABTIM3-hum21 wasdetermined by x-ray crystallography studies. ABTIM3-hum21 differs byonly one amino acid in the heavy chain CDR2 from the other humanizedanti-TIM3 antibodies described herein, and this different amino acid(Gln55) is far away (>6 Å) from the epitope and thus would not beexpected to change antigen binding. Two different competition assayswere performed to compare epitope binding between ABTIM3-hum21 and twoother humanized anti-TIM3 antibodies, ABTIM3-hum03 and ABTIM3-hum11. Theresults of both competition assays show that both ABTIM3-hum04 andABTIM3-hum11 effectively compete with ABTIM3-hum03 for binding to TIM3,thus demonstrating that ABTIM3-hum03 and ABTIM3-hum11 also bind to asimilar epitope as ABTIM3-hum21, e.g., the epitope as described herein.

11.1 Flow Cytometry Competition Assay

K_(D) of ABTMI3-hum21 was determined by labeling ABTIM3-hum21 withphycoerythrin, incubated with 300.19 hTIM-3 expressing cells, and abinding curve was established to determine a K_(D) of 2.15.

Titrated concentrations of unlabelled hIgG1(isotype control),ABTIM3-hum21 (positive control), ABTIM3-hum11 or ABTIM3-hum03 were mixedwith ABTIM3-hum21 at its K_(D) and incubated with 300.19 hTIM-3expressing cells at 4° C. for 3 hours. Cells were washed twice and runon an LSRFortessa flow cytometer. Data was analyzed in FlowJo and MFI(PE) values were plotted and graphed in GraphPad (Prism) software. Theexperiment was performed twice.

The results of the competition assay demonstrate that ABTIM3-hum11 andABTIM3-hum03 (but not isotype control) both competed with ABTIM3-hum21for binding with human TIM3 expressed on the 300.19 cells (FIG. 20).K_(D) for ABTIM3-hum11 and ABTIM3-hum03 was calculated from the bindingcurves; the calculated K_(D) for ABTIM3-hum11 was 2.276 nM and thecalculated K_(D) for ABTIM3-hum03 was 2.413 nM. These resultsdemonstrate that ABTIM3-hum11 and ABTIM3-hum03 bind to a similar or thesame epitope as ABTIM3-hum21.

11.2 Biacore Competition Assay

hTIM-3/his antigen was captured by immobilized anti-His antibody(RU10000) on a CM5 chip. The first antibody was injected to reachsaturation (>90%). Then the second antibody was injected to assesswhether a second binding event occurs. Occurrence of a second bindingevent indicates that the two tested antibodies have different epitopes.Lack of a second binding event, indicates that the two antibodies mayrecognize and bind to the same epitope. Control assays were run where atest antibody was run with human IgG1 isotype control, or where the testantibody was run as the first and second antibody (e.g., self-selfcycle) to observe the baseline of a binding event. Table 14 summarizesthe Biacore cycles run and indicates which antibodies were used as thefirst and second antibody in each cycle.

TABLE 14 Summary of Biacore competition assay cycles Cycles 1^(st)Antibody 2^(nd) Antibody 1 huIgG1 huIgG1 2 huIgG1 ABTIM3-hum21 3 huIgG1ABTIM3-hum03 4 huIgG1 ABTIM3-hum11 5 ABTIM3-hum21 huIgG1 6 ABTIM3-hum21ABTIM3-hum21 7 ABTIM3-hum21 ABTIM3-hum03 8 ABTIM3-hum21 ABTIM3-hum11 9ABTIM3-hum03 huIgG1 10 ABTIM3-hum03 ABTIM3-hum21 11 ABTIM3-hum03ABTIM3-hum03 12 ABTIM3-hum03 ABTIM3-hum11 13 ABTIM3-hum11 huIgG1 14ABTIM3-hum11 ABTIM3-hum21 15 ABTIM3-hum11 ABTIM3-hum03 16 ABTIM3-hum11ABTIM3-hum11

Detection of the baseline and first and second binding events arerecorded as RU (resonance units) and can be presented in a sensogram. Atypical sensogram is shown in FIG. 21, where a binding event is shownafter the 1^(st) antibody injection. After a wash, the second antibodyis injected and a second binding may be detected. Significant changes inRU indicate a binding event. A summary of the changes in RU detectedfrom the 1^(st) and 2^(nd) antibody injections from the Biacorecompetition assay is shown in Table 15.

TABLE 15 Summary of results from Biacore competition assay 2^(nd)Antibody Injection ABTIM3- ABTIM3- ABTIM3- 1^(st) Antibody InjectionhuIgG1 hum21 hum03 hum11 huIgG1 0.27 3.6 88.2 86.3 83.2 ABTIM3-hum2195.85 4.5 6.6 7.6 8.1 ABTIM3-hum03 93.33 4.5 6.9 7.3 8.5 ABTIM3-hum1193.48 3.8 NA¹ 5.3 7.2 ¹No value was calculated from the sensogram, dueto an unknown fluid problem.

The results shown above demonstrate that injection of ABTIM3-hum21,ABTIM3-hum03, and ABTIM3-hum11 during the first antibody injectionresults in a binding event. Injection of ABTIM3-hum21, ABTIM3-hum03, andABTIM3-hum11 as the second antibody after injection is human IgG1control antibody results in a second binding event. However, injectionof any of the anti-TIM3 antibodies tested here as the first and secondantibodies did not result in a second binding event, demonstrating thatfor each pair of 1^(st) and 2^(nd) antibodies tested, there wascompetition for binding to the same TIM3 epitope. These results indicatethat ABTIM3-hum21, ABTIM3-hum03, and ABTIM3-hum11 bind to a similar orthe same epitope on human TIM3.

Example 12 Pharmacokinetic Properties of ABTIM3-Hum11

Various pharmacokinetic properties of ABTIM3-hum11 were assessed inmouse and rat models. ABTIM3-hum11 was injected intravenously into miceat varying doses, 1 mg/kg, 3 mg/kg, and 10 mg/kg. Blood samples wereobtained at various timepoints between 0 and 672 hours (0-28 days). 10mg/kg ABTIM3-hum11 was injected intravenously into rats, and bloodsamples were obtained at various time points from 0-400 hours (0-16days). The concentration of ABTIM3-hum11 present in the serum wasdetermined (FIGS. 23A and 23B). The results showed that ABTIM3-hum11 isstable in both mouse and rat serum. Table 16 shows additionalpharmacokinetic properties determined, including halflife (T1/2), peakserum concentration (Cmax), AUC up to the last measurable concentration(AUClast), and AUC as extrapolated to infinity (AUCinf).

TABLE 16 Summary of pharmacokinetic properties of ABTIM3-hum11 Dose T ½Cmax AUClast AUCinf Species (mg/kg) (hr) (μg/mL) (hr*μg/mL) (hr*μg/mL)Mouse  1 N 3 3 3 3 Mean 142.3 17.3 1507.8 1571.4 STD 96.9 0.7 337.9439.5  3 N 3 3 3 3 Mean 266.1 37.2 4617.9 5369.0 STD 73.1 2.3 2109.82496.1 10 N 3 3 3 3 Mean 254.9 147.5 23906.5 28621.7 STD 39.2 13.24369.8 6314.1 Rat 10 N 3 3 3 3 Mean 400.8 243.4 26032.3 53767.1 STD 75.919.1 895.8 5362.6

In a toxicity study, three naïve mice were administered a single dose byintravenous injection at 1 mg/kg, 3 mg/kg, or 10 mg/kg of ABTIM3-huml 1.After 28 days, no adverse events were observed, indicating that ABTIM3antibody is tolerable in mouse models.

INCORPORATION BY REFERENCE

All publications, patents, and Accession numbers mentioned herein arehereby incorporated by reference in their entirety as if each individualpublication or patent was specifically and individually indicated to beincorporated by reference.

EQUIVALENTS

While specific embodiments of the compositions and methods herein havebeen discussed, the above specification is illustrative and notrestrictive. Many variations of the invention will become apparent tothose skilled in the art upon review of this specification and theclaims below. The full scope of the invention should be determined byreference to the claims, along with their full scope of equivalents, andthe specification, along with such variations.

What is claimed is:
 1. A nucleic acid molecule comprising a nucleotidesequence selected from: a nucleotide sequence that encodes a heavy chainvariable region (VH), a nucleotide sequence that encodes a light chainvariable region (VL), or a nucleotide sequence that encodes both a VHand a VL of an antibody molecule that binds to human T-cellimmunoglobulin domain and mucin domain 3 (TIM-3), wherein the antibodymolecule comprises: (a) a VH comprising a VHCDR1 amino acid sequence ofSEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQ ID NO: 10; and aVHCDR3 amino acid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1amino acid sequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence ofSEQ ID NO: 13, and a VLCDR3 amino acid sequence of SEQ ID NO: 14; (b) aVH comprising a VHCDR1 amino acid sequence of SEQ ID NO: 3; a VHCDR2amino acid sequence of SEQ ID NO: 4; and a VHCDR3 amino acid sequence ofSEQ ID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ IDNO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7, and a VLCDR3 aminoacid sequence of SEQ ID NO: 8; (c) a VH comprising a VHCDR1 amino acidsequence of SEQ ID NO: 9; a VHCDR2 amino acid sequence of SEQ ID NO: 25;and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and a VL comprising aVLCDR1 amino acid sequence of SEQ ID NO: 12, a VLCDR2 amino acidsequence of SEQ ID NO: 13, and a VLCDR3 amino acid sequence of SEQ IDNO: 14; (d) a VH comprising a VHCDR1 amino acid sequence of SEQ ID NO:3; a VHCDR2 amino acid sequence of SEQ ID NO: 24; and a VHCDR3 aminoacid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO: 8; (e) a VH comprising aVHCDR1 amino acid sequence of SEQ ID NO: 9; a VHCDR2 amino acid sequenceof SEQ ID NO: 31; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; anda VL comprising a VLCDR1 amino acid sequence of SEQ ID NO: 12, a VLCDR2amino acid sequence of SEQ ID NO: 13, and a VLCDR3 amino acid sequenceof SEQ ID NO: 14; or (f) a VH comprising a VHCDR1 amino acid sequence ofSEQ ID NO: 3; a VHCDR2 amino acid sequence of SEQ ID NO: 30; and aVHCDR3 amino acid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1amino acid sequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQID NO: 7, and a VLCDR3 amino acid sequence of SEQ ID NO:
 8. 2. Thenucleic acid molecule of claim 1, wherein the nucleotide sequencesencoding the VH and VL of the antibody molecule are in the same nucleicacid molecule.
 3. The nucleic acid molecule of claim 1, wherein thenucleotide sequences encoding the VH and VL of the antibody molecule arein separate nucleic acid molecules.
 4. The nucleic acid molecule ofclaim 1, wherein the antibody molecule comprises a VH comprising aVHCDR1 amino acid sequence of SEQ ID NO: 9; a VHCDR2 amino acid sequenceof SEQ ID NO: 10; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; anda VL comprising a VLCDR1 amino acid sequence of SEQ ID NO: 12, a VLCDR2amino acid sequence of SEQ ID NO: 13, and a VLCDR3 amino acid sequenceof SEQ ID NO:
 14. 5. The nucleic acid molecule of claim 1, wherein theantibody molecule comprises a VH comprising a VHCDR1 amino acid sequenceof SEQ ID NO: 3; a VHCDR2 amino acid sequence of SEQ ID NO: 4; and aVHCDR3 amino acid sequence of SEQ ID NO: 5; and a VL comprising a VLCDR1amino acid sequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQID NO: 7, and a VLCDR3 amino acid sequence of SEQ ID NO:
 8. 6. Thenucleic acid molecule of claim 1, wherein the antibody moleculecomprises a VH comprising a VHCDR1 amino acid sequence of SEQ ID NO: 9;a VHCDR2 amino acid sequence of SEQ ID NO: 25; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 12, a VLCDR2 amino acid sequence of SEQ ID NO:13, and a VLCDR3 amino acid sequence of SEQ ID NO:
 14. 7. The nucleicacid molecule of claim 1, wherein the antibody molecule comprising a VHcomprising a VH comprising a VHCDR1 amino acid sequence of SEQ ID NO: 3;a VHCDR2 amino acid sequence of SEQ ID NO: 24; and a VHCDR3 amino acidsequence of SEQ ID NO: 5; and a VL comprising a VLCDR1 amino acidsequence of SEQ ID NO: 6, a VLCDR2 amino acid sequence of SEQ ID NO: 7,and a VLCDR3 amino acid sequence of SEQ ID NO:
 8. 8. The nucleic acidmolecule of claim 1 wherein the antibody molecule comprising a VHcomprising a VHCDR1 amino acid sequence of SEQ ID NO: 9; a VHCDR2 aminoacid sequence of SEQ ID NO: 31; and a VHCDR3 amino acid sequence of SEQID NO: 5; and a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO:12, a VLCDR2 amino acid sequence of SEQ ID NO: 13, and a VLCDR3 aminoacid sequence of SEQ ID NO:
 14. 9. The nucleic acid molecule of claim 1,wherein the antibody molecule comprises a VH comprising a VHCDR1 aminoacid sequence of SEQ ID NO: 3; a VHCDR2 amino acid sequence of SEQ IDNO: 30; and a VHCDR3 amino acid sequence of SEQ ID NO: 5; and a VLcomprising a VLCDR1 amino acid sequence of SEQ ID NO: 6, a VLCDR2 aminoacid sequence of SEQ ID NO: 7, and a VLCDR3 amino acid sequence of SEQID NO:
 8. 10. The nucleic acid molecule of claim 1, wherein thenucleotide sequence encodes a VH comprising an amino acid sequence atleast 85% identical to any of SEQ ID NOs: 1, 16, 26, 32, 36, 44, 48, 52,60, 68, 72, 76, 80, 84, 92, or
 100. 11. The nucleic acid molecule ofclaim 1, wherein the nucleotide sequence encodes a VH comprising theamino acid sequence of SEQ ID NO: 1, 16, 26, 32, 36, 44, 48, 52, 60, 68,72, 76, 80, 84, 92, or
 100. 12. The nucleic acid molecule of claim 1,wherein the nucleotide sequence encodes a VL comprising an amino acidsequence at least 85% identical to any of SEQ ID NOs: 2, 20, 40, 56, 64,88, 96, or
 104. 13. The nucleic acid molecule of claim 1, wherein thenucleotide sequence encodes a VL comprising the amino acid sequence ofSEQ ID NO: 2, 20, 40, 56, 64, 88, 96, or
 104. 14. The nucleic acidmolecule of claim 1, wherein the nucleotide sequence comprises anucleotide sequence at least 85% identical to any of SEQ ID NOs: 11, 17,29, 33, 37, 45, 49, 53, 61, 69, 73, 77, 81, 85, 93, 101, 115, or 120.15. The nucleic acid molecule of claim 1, wherein the nucleotidesequence comprises the nucleotide sequence of SEQ ID NO: 11, 17, 29, 33,37, 45, 49, 53, 61, 69, 73, 77, 81, 85, 93, 101, 115, or
 120. 16. Thenucleic acid molecule of claim 1, wherein the nucleotide sequencecomprises a nucleotide sequence at least 85% identical to any of SEQ IDNOs: 15, 21, 41, 57, 65, 89, 97, 105, 118, 123, 125, or
 127. 17. Thenucleic acid molecule of claim 1, wherein the nucleotide sequencecomprises the nucleotide sequence of SEQ ID NO: 15, 21, 41, 57, 65, 89,97, 105, 118, 123, 125, or
 127. 18. The nucleic acid molecule of claim1, wherein the nucleotide sequence encodes: (a) a VH comprising theamino acid sequence of SEQ ID NO: 1 and a VL comprising the amino acidsequence of SEQ ID NO: 2; (b) a VH comprising the amino acid sequence ofSEQ ID NO: 16 and a VL comprising the amino acid sequence of SEQ ID NO:20; (c) a VH comprising the amino acid sequence of SEQ ID NO: 26 and aVL comprising the amino acid sequence of SEQ ID NO: 20; (d) a VHcomprising the amino acid sequence of SEQ ID NO: 32 and a VL comprisingthe amino acid sequence of SEQ ID NO: 20; (e) a VH comprising the aminoacid sequence of SEQ ID NO: 36 and a VL comprising the amino acidsequence of SEQ ID NO: 40; (f) a VH comprising the amino acid sequenceof SEQ ID NO: 44 and a VL comprising the amino acid sequence of SEQ IDNO: 40; (g) a VH comprising the amino acid sequence of SEQ ID NO: 48 anda VL comprising the amino acid sequence of SEQ ID NO: 40; (h) a VHcomprising the amino acid sequence of SEQ ID NO: 36 and a VL comprisingthe amino acid sequence of SEQ ID NO: 20; (i) a VH comprising the aminoacid sequence of SEQ ID NO: 16 and a VL comprising the amino acidsequence of SEQ ID NO: 40; (j) a VH comprising the amino acid sequenceof SEQ ID NO: 52 and a VL comprising the amino acid sequence of SEQ IDNO: 56; (k) a VH comprising the amino acid sequence of SEQ ID NO: 60 anda VL comprising the amino acid sequence of SEQ ID NO: 56; (l) a VHcomprising the amino acid sequence of SEQ ID NO: 52 and a VL comprisingthe amino acid sequence of SEQ ID NO: 64; (m) a VH comprising the aminoacid sequence of SEQ ID NO: 60 and a VL comprising the amino acidsequence of SEQ ID NO: 64; (n) a VH comprising the amino acid sequenceof SEQ ID NO: 68 and a VL comprising the amino acid sequence of SEQ IDNO: 64; (o) a VH comprising the amino acid sequence of SEQ ID NO: 72 anda VL comprising the amino acid sequence of SEQ ID NO: 64; (p) a VHcomprising the amino acid sequence of SEQ ID NO: 76 and a VL comprisingthe amino acid sequence of SEQ ID NO: 56; (q) a VH comprising the aminoacid sequence of SEQ ID NO: 80 and a VL comprising the amino acidsequence of SEQ ID NO: 56; (r) a VH comprising the amino acid sequenceof SEQ ID NO: 68 and a VL comprising the amino acid sequence of SEQ IDNO: 56; (s) a VH comprising the amino acid sequence of SEQ ID NO: 72 anda VL comprising the amino acid sequence of SEQ ID NO: 56; (t) a VHcomprising the amino acid sequence of SEQ ID NO: 76 and a VL comprisingthe amino acid sequence of SEQ ID NO: 64; (u) a VH comprising the aminoacid sequence of SEQ ID NO: 80 and a VL comprising the amino acidsequence of SEQ ID NO: 64; (v) a VH comprising the amino acid sequenceof SEQ ID NO: 84 and a VL comprising the amino acid sequence of SEQ IDNO: 88; (w) a VH comprising the amino acid sequence of SEQ ID NO: 92 anda VL comprising the amino acid sequence of SEQ ID NO: 96; or (x) a VHcomprising the amino acid sequence of SEQ ID NO: 100 and a VL comprisingthe amino acid sequence of SEQ ID NO:
 104. 19. The nucleic acid moleculeof claim 1, which comprises: (a) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 11 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 15; (b) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 17 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 21; (c) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 27 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 21; (d) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 33 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 21; (e) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 37 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 41; (f) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 45 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 41; (g) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 49 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 41; (h) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 115 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 118; (i) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 120 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 123; (j) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 53 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 57; (k) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 61 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 57; (l) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 53 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 65; (m) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 61 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 65; (n) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 69 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 125; (o) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 73 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 125; (p) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 77 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 127; (q) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 81 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 127; (r) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 69 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 127; (s) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 73 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 127; (t) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 77 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 125; (u) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 81 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 125; (v) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 85 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO: 89; (w) a first nucleotide sequence comprisingthe nucleotide sequence of SEQ ID NO: 93 and a second nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 97; or (x) afirst nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 101 and a second nucleotide sequence comprising the nucleotidesequence of SEQ ID NO:
 105. 20. The nucleic acid molecule of claim 1,which encodes a heavy chain comprising an amino acid sequence at least85% identical to any of SEQ ID NOs: 18, 28, 34, 38, 46, 50, 54, 62, 70,74, 78, 82, 86, 94, 102, 116, or
 121. 21. The nucleic acid molecule ofclaim 1, which encodes a heavy chain comprising the amino acid sequenceof SEQ ID NO: 18, 28, 34, 38, 46, 50, 54, 62, 70, 74, 78, 82, 86, 94,102, 116, or
 121. 22. The nucleic acid molecule of claim 1, whichencodes a light chain comprising an amino acid sequence at least 85%identical to any of SEQ ID NOs: 22, 42, 58, 66, 90, 98, or
 106. 23. Thenucleic acid molecule of claim 1, which encodes a light chain comprisingthe amino acid sequence of SEQ ID NO: 22, 42, 58, 66, 90, 98, or 106.24. The nucleic acid molecule of claim 1, which comprises a heavy chainnucleotide sequence at least 85% identical to any of SEQ ID NOs: 19, 29,35, 39, 47, 51, 55, 63, 71, 75, 79, 83, 87, 95, 103, 117, or
 122. 25.The nucleic acid molecule of claim 1, which comprises a heavy chainnucleotide sequence of SEQ ID NO: 19, 29, 35, 39, 47, 51, 55, 63, 71,75, 79, 83, 87, 95, 103, 117, or
 1227. 26. The nucleic acid molecule ofclaim 1, which comprises a light chain nucleotide sequence at least 85%identical to any of SEQ ID NOs: 23, 43, 59, 67, 91, 99, 107, 119, 124,126, or
 128. 27. The nucleic acid molecule of claim 1, which comprises alight chain nucleotide sequence of SEQ ID NO: 23, 43, 59, 67, 91, 99,107, 119, 124, 126, or
 128. 28. The nucleic acid molecule of claim 1,which encodes: (a) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 18 and a light chain comprising the amino acid sequence ofSEQ ID NO: 22; (b) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 28 and a light chain comprising the amino acid sequence ofSEQ ID NO: 22; (c) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 34 and a light chain comprising the amino acid sequence ofSEQ ID NO: 22; (d) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 38 and a light chain comprising the amino acid sequence ofSEQ ID NO: 42; (e) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 46 and a light chain comprising the amino acid sequence ofSEQ ID NO: 42; (f) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 50 and a light chain comprising the amino acid sequence ofSEQ ID NO: 42; (g) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 116 and a light chain comprising the amino acid sequence ofSEQ ID NO: 22; (h) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 121 and a light chain comprising the amino acid sequence ofSEQ ID NO: 42; (i) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 54 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (j) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 62 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (k) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 54 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (l) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 62 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (m) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 70 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (n) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 74 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (o) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 78 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (p) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 82 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (q) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 70 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (r) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 74 and a light chain comprising the amino acid sequence ofSEQ ID NO: 58; (s) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 78 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (t) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 82 and a light chain comprising the amino acid sequence ofSEQ ID NO: 66; (u) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 86 and a light chain comprising the amino acid sequence ofSEQ ID NO: 90; (v) a heavy chain comprising the amino acid sequence ofSEQ ID NO: 94 and a light chain comprising the amino acid sequence ofSEQ ID NO: 98; or (w) a heavy chain comprising the amino acid sequenceof SEQ ID NO: 102 and a light chain comprising the amino acid sequenceof SEQ ID NO:
 106. 29. The nucleic acid molecule of claim 1, whichcomprises: (a) a heavy chain nucleotide sequence of SEQ ID NO: 19 and alight chain nucleotide sequence of SEQ ID NO: 23; (b) a heavy chainnucleotide sequence of SEQ ID NO: 29 and a light chain nucleotidesequence of SEQ ID NO: 23; (c) a heavy chain nucleotide sequence of SEQID NO: 35 and a light chain nucleotide sequence of SEQ ID NO: 23; (d) aheavy chain nucleotide sequence of SEQ ID NO: 39 and a light chainnucleotide sequence of SEQ ID NO: 43; (e) a heavy chain nucleotidesequence of SEQ ID NO: 47 and a light chain nucleotide sequence of SEQID NO: 43; (f) a heavy chain nucleotide sequence of SEQ ID NO: 51 and alight chain nucleotide sequence of SEQ ID NO: 43; (g) a heavy chainnucleotide sequence of SEQ ID NO: 117 and a light chain nucleotidesequence of SEQ ID NO: 119; (h) a heavy chain nucleotide sequence of SEQID NO: 122 and a light chain nucleotide sequence of SEQ ID NO: 124; (i)a heavy chain nucleotide sequence of SEQ ID NO: 55 and a light chainnucleotide sequence of SEQ ID NO: 59; (j) a heavy chain nucleotidesequence of SEQ ID NO: 63 and a light chain nucleotide sequence of SEQID NO: 59; (k) a heavy chain nucleotide sequence of SEQ ID NO: 55 and alight chain nucleotide sequence of SEQ ID NO: 67; (l) a heavy chainnucleotide sequence of SEQ ID NO: 63 and a light chain nucleotidesequence of SEQ ID NO: 67; (m) a heavy chain nucleotide sequence of SEQID NO: 71 and a light chain nucleotide sequence of SEQ ID NO: 126; (n) aheavy chain nucleotide sequence of SEQ ID NO: 75 and a light chainnucleotide sequence of SEQ ID NO: 126; (o) a heavy chain nucleotidesequence of SEQ ID NO: 79 and a light chain nucleotide sequence of SEQID NO: 128; (p) a heavy chain nucleotide sequence of SEQ ID NO: 83 and alight chain nucleotide sequence of SEQ ID NO: 128; (q) a heavy chainnucleotide sequence of SEQ ID NO: 71 and a light chain nucleotidesequence of SEQ ID NO: 128; (r) a heavy chain nucleotide sequence of SEQID NO: 75 and a light chain nucleotide sequence of SEQ ID NO: 128; (s) aheavy chain nucleotide sequence of SEQ ID NO: 79 and a light chainnucleotide sequence of SEQ ID NO: 126; (t) a heavy chain nucleotidesequence of SEQ ID NO: 83 and a light chain nucleotide sequence of SEQID NO: 126; (u) a heavy chain nucleotide sequence of SEQ ID NO: 87 and alight chain nucleotide sequence of SEQ ID NO: 91; (v) a heavy chainnucleotide sequence of SEQ ID NO: 95 and a light chain nucleotidesequence of SEQ ID NO: 99; or (w) a heavy chain nucleotide sequence ofSEQ ID NO: 103 and a light chain nucleotide sequence of SEQ ID NO: 107.30. The nucleic acid molecule of claim 1, which encodes a Fab, F(ab′)2,Fv, or a single chain Fv fragment (scFv).
 31. The nucleic acid moleculeof claim 1, further comprising a nucleotide sequence encoding a heavychain constant region of IgG1, IgG2, IgG3, or IgG4.
 32. The nucleic acidmolecule of claim 1, further comprising a nucleotide sequence encoding alight chain constant region of kappa or lambda.
 33. The nucleic acidmolecule of claim 1, further comprising a nucleotide sequence encoding ahuman IgG4 heavy chain constant region with a Serine to Proline mutationat position 108 of SEQ ID NO: 108 or 110 and a nucleotide sequenceencoding a kappa light chain constant region.
 34. The nucleic acidmolecule of claim 1, further comprising a nucleotide sequence encoding ahuman IgG1 heavy chain constant region with an Asparagine to Alaninemutation at position 180 of SEQ ID NO: 112 and a nucleotide sequenceencoding a kappa light chain constant region.
 35. The nucleic acidmolecule of claim 1, further comprising a nucleotide sequence encoding ahuman IgG1 heavy chain constant region with an Aspartate to Alaninemutation at position 148 of SEQ ID NO: 113 and Proline to Alaninemutation at position 212 of SEQ ID NO: 113, and a nucleotide sequenceencoding a kappa light chain constant region.
 36. The nucleic acidmolecule of claim 1, further comprising a nucleotide sequence encoding ahuman IgG1 heavy chain constant region with a Leucine to Alaninemutation at position 117 of SEQ ID NO: 114 and Leucine to Alaninemutation at position 118 of SEQ ID NO: 114, and a nucleotide sequenceencoding a kappa light chain constant region.
 37. The nucleic acidmolecule of claim 1, wherein the nucleotide sequence encodes a VHcomprising the amino acid sequence of SEQ ID NO: 32 and a VL comprisingthe amino acid sequence of SEQ ID NO:
 20. 38. The nucleic acid moleculeof claim 1, wherein the nucleotide sequence encodes a VH comprising theamino acid sequence of SEQ ID NO: 52 and a VL comprising the amino acidsequence of SEQ ID NO:
 64. 39. The nucleic acid molecule of claim 1,comprising the nucleotide sequence of SEQ ID NO: 33 and SEQ ID NO: 21.40. The nucleic acid molecule of claim 1, comprising the nucleotidesequence of SEQ ID NO: 53 and SEQ ID NO:
 65. 41. The nucleic acidmolecule of claim 1, wherein the nucleotide sequence encodes a heavychain comprising the amino acid sequence of SEQ ID NO: 34 and a lightchain comprising the amino acid sequence of SEQ ID NO:
 22. 42. Thenucleic acid molecule of claim 1, wherein the nucleotide sequenceencodes a heavy chain comprising the amino acid sequence of SEQ ID NO:54 and a light chain comprising the amino acid sequence of SEQ ID NO:66.
 43. The nucleic acid molecule of claim 1, comprising a heavy chainnucleotide sequence of SEQ ID NO: 35 and a light chain nucleotidesequence of SEQ ID NO:
 23. 44. The nucleic acid molecule of claim 1,comprising a heavy chain nucleotide sequence of SEQ ID NO: 55 and alight chain nucleotide sequence of SEQ ID NO:
 67. 45. An expressionvector comprising the nucleic acid molecule of claim
 1. 46. An isolatedhost cell comprising the nucleic acid molecule of claim
 1. 47. A methodof producing an antibody molecule, comprising culturing the host cell ofclaim 46 under conditions suitable for gene expression.
 48. A nucleicacid molecule comprising a first nucleotide sequence encoding a VH and asecond nucleotide sequence encoding a VL, of an antibody molecule thatbinds to human TIM-3, wherein the VH comprises the amino acid sequenceof SEQ ID NO: 32 and the VL comprises the amino acid sequence of SEQ IDNO:
 20. 49. An expression vector comprising the nucleic acid molecule ofclaim
 48. 50. An isolated host cell comprising the nucleic acid moleculeof claim
 48. 51. A method of producing an antibody molecule or fragmentthereof, comprising culturing the host cell of claim 50 under conditionssuitable for gene expression, thereby producing the antibody molecule orfragment thereof.
 52. A nucleic acid molecule comprising a firstnucleotide sequence encoding a VH and a second nucleotide sequenceencoding a VL, of an antibody molecule that binds to human TIM-3,wherein the VH comprises the amino acid sequence of SEQ ID NO: 52 andthe VL comprises the amino acid sequence of SEQ ID NO:
 64. 53. Anexpression vector comprising the nucleic acid molecule of claim
 52. 54.An isolated host cell comprising the nucleic acid molecule of claim 52.55. A method of producing an antibody molecule or fragment thereof,comprising culturing the host cell of claim 54 under conditions suitablefor gene expression, thereby producing the antibody molecule or fragmentthereof.
 56. A nucleic acid molecule comprising a first nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 33 and asecond nucleotide sequence comprising the nucleotide sequence of SEQ IDNO:
 21. 57. A nucleic acid molecule comprising a first nucleotidesequence comprising the nucleotide sequence of SEQ ID NO: 53 and asecond nucleotide sequence comprising the nucleotide sequence of SEQ IDNO: 65.